STEP 1 -

Thymine


STEP 1 -

STEP 2
The second step is removal of the DMT protecting group by treatment with dichloroacetic acid in $\mathrm{CH}_{2} \mathrm{Cl}_{2}$. The reaction occurs by an $\mathrm{S}_{\mathrm{N}} 1$ mechanism and proceeds rapidly because of the stability of the tertiary, benzylic dimethoxytrityl cation.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1026.jpg?height=343&width=871&top_left_y=1517&top_left_x=622)


STEP 1 -

STEP 3
The third step is the coupling of the polymer-bonded deoxynucleoside with a protected deoxynucleoside containing a phosphoramidite group $\left[\mathrm{R}_{2} \mathrm{NP}(\mathrm{OR})_{2}\right]$ at its $3^{\prime}$ position. The coupling reaction takes place in the polar aprotic solvent acetonitrile, requires catalysis by the heterocyclic amine tetrazole, and yields a phosphite, $\mathrm{P}(\mathrm{OR})_{3}$, as product. Note that one of the phosphorus oxygen atoms is protected by a $\beta$-cyanoethyl group, $-\mathrm{OCH}_{2} \mathrm{CH}_{2} \mathrm{C} \equiv \mathrm{N}$. The coupling step takes place with better than $99 \%$ yield.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1027.jpg?height=421&width=389&top_left_y=332&top_left_x=288)
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1027.jpg?height=605&width=583&top_left_y=196&top_left_x=1248)


STEP 4 -

With the coupling accomplished, the phosphite product is oxidized to a phosphate by treatment with iodine in aqueous tetrahydrofuran in the presence of 2,6-dimethylpyridine. The cycle of (1) deprotection, (2) coupling, and (3) oxidation is then repeated until an oligonucleotide chain of the desired sequence has been constructed.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1027.jpg?height=605&width=1494&top_left_y=996&top_left_x=310)


STEP 4 -

STEP 5
The final step is removal of all protecting groups and cleavage of the ester bond holding the DNA to the silica. All these reactions are done at the same time by treatment with aqueous $\mathrm{NH}_{3}$. Purification by electrophoresis then yields the synthetic DNA.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1028.jpg?height=801&width=936&top_left_y=185&top_left_x=589)


STEP 4 -

PROBLEM p-Dimethoxytrityl (DMT) ethers are easily cleaved by mild acid treatment. Show the mechanism of 28-11 the cleavage reaction.


STEP 4 -

PROBLEM Propose a mechanism to account for cleavage of the $\beta$-cyanoethyl protecting group from the 28-12 phosphate groups on treatment with aqueous ammonia. (Acrylonitrile, $\mathrm{H}_{2} \mathrm{C}=\mathrm{CHCN}$, is a byproduct.) What kind of reaction is occurring?


STEP 4 - 28.8 The Polymerase Chain Reaction

It often happens that only a tiny amount of DNA can be obtained directly, as might occur at a crime scene, so methods for obtaining larger amounts are sometimes needed to carry out sequencing and characterization. The invention of the polymerase chain reaction (PCR) by Kary Mullis in 1986 has been described as being to genes what Gutenberg's invention of the printing press was to the written word. Just as the printing press produces multiple copies of a book, PCR produces multiple copies of a given DNA sequence. Starting from less than 1 picogram ( pg ) of DNA with a chain length of 10,000 nucleotides ( $1 \mathrm{pg}=10^{-12} \mathrm{~g}$; about $10^{5}$ molecules), PCR makes it possible to obtain several micrograms ( $1 \mu \mathrm{~g}=10^{-6} \mathrm{~g}$; about $10^{11}$ molecules) in just a few hours.


The key to the polymerase chain reaction is Taq DNA polymerase, a heat-stable enzyme isolated from the thermophilic bacterium Thermus aquaticus found in a hot spring in Yellowstone National Park. Taq polymerase is able to take a single strand of DNA having a short, primer segment of complementary chain at one end and then finish constructing the entire complementary strand. The overall process takes three steps, as shown in FIGURE 28.10. More recently, improved heat-stable DNA polymerases have become available, including Vent polymerase and Pfu polymerase, both isolated from bacteria growing near geothermal vents in the ocean floor. The error rate of both enzymes is substantially less than that of Taq.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1029.jpg?height=998&width=1215&top_left_y=192&top_left_x=452)


FIGURE 28.10 The polymerase chain reaction. Details are explained in the text.
STEP 1
The double-stranded DNA to be amplified is heated in the presence of Taq polymerase, $\mathrm{Mg}^{2+}$ ion, the four deoxynucleotide triphosphate monomers (dNTPs), and a large excess of two short oligonucleotide primers of about 20 bases each. Each primer is complementary to the sequence at the end of one of the target DNA segments. At a temperature of $95^{\circ} \mathrm{C}$, double-stranded DNA denatures, spontaneously breaking apart into two single strands.


STEP 2
The temperature is lowered to between 37 and $50^{\circ} \mathrm{C}$, allowing the primers, because of their relatively high concentration, to anneal by hydrogen-bonding to their complementary sequence at the end of each target strand.


STEP 3
The temperature is then raised to $72^{\circ} \mathrm{C}$, and Taq polymerase catalyzes the addition of further nucleotides to the two primed DNA strands. When replication of each strand is complete, two copies of the original DNA now exist. Repeating the denature-anneal-synthesize cycle a second time yields four DNA copies, repeating a third time yields eight copies, and so on, in an exponential series.


PCR has been automated, and 30 or so cycles can be carried out in an hour, resulting in a theoretical amplification factor of $2^{30}\left(\sim 10^{9}\right)$. In practice, however, the efficiency of each cycle is less than $100 \%$, and an experimental amplification of about $10^{6}$ to $10^{8}$ is routinely achieved for 30 cycles.


DNA Fingerprinting -

The invention of DNA sequencing has affected society in many ways, few more dramatic than those stemming from the development of DNA fingerprinting. DNA fingerprinting arose from the discovery in 1984 that human genes contain short, repeating sequences of noncoding DNA, called short tandem repeat (STR) loci. Furthermore, the STR loci are slightly different for everyone except identical twins. By sequencing these loci, a pattern unique to each person can be obtained.


DNA Fingerprinting -

Perhaps the most common and well-publicized use of DNA fingerprinting is that carried out by crime laboratories to link suspects to biological evidence-blood, hair follicles, skin, or semen-found at a crime scene. Many thousands of court cases have been decided based on DNA evidence.


DNA Fingerprinting -

For use in criminal cases, forensic laboratories in the United States have agreed on 13 core STR loci that are most accurate for the identification of an individual. Based on these 13 loci, a Combined DNA Index System (CODIS) has been established to serve as a registry of convicted offenders. When a DNA sample is obtained from a crime scene, the sample is subjected to cleavage with restriction endonucleases to cut out fragments containing the STR loci, the fragments are amplified using the polymerase chain reaction, and the sequences of the fragments are determined.


DNA Fingerprinting -

If the profile of sequences from a known individual and the profile from DNA obtained at a crime scene match, the probability is approximately 82 billion to 1 that the DNA is from the same individual. In paternity cases, where the DNA of male parent and offspring are related but not fully identical, the identity of the male parent can be established with a probability of around 100,000 to 1 . Even after several generations, paternity can still be inferred from DNA analysis of the $Y$ chromosome of direct male-line descendants. The most well-known such case is that of Thomas Jefferson, one of the nation's founders, who likely impregnated enslaved person Sally Hemings. Although Jefferson himself has no male-line descendants, DNA analysis of the male-line descendants of Jefferson's paternal uncle contained the same Y chromosome as a male-line descendant of Eston Hemings, the youngest son of Sally Hemings. Thus, a mixing of the two genomes is clear, although the male individual responsible for that mixing can't be identified with $100 \%$ certainty.


DNA Fingerprinting -

Among its many other applications, DNA fingerprinting is widely used for the diagnosis of genetic disorders, both prenatally and in newborns. Cystic fibrosis, hemophilia, Huntington's disease, Tay-Sachs disease, sickle cell anemia, and thalassemia are among the many diseases that can be detected, enabling early treatment of an affected child. Furthermore, by studying the DNA fingerprints of relatives with a history of a particular disorder, it's possible to identify DNA patterns associated with the disease and perhaps obtain clues for an eventual cure. In addition, the U.S. Department of Defense requires blood and saliva samples from all military personnel. The samples are stored, and DNA is extracted if the need for identification of a casualty arises.


Key Terms -

- anticodon
- antisense strand
- codon
- deoxyribonucleic acid (DNA)
- double helix
- 3' end
- 5' end
- messenger RNA (mRNA)
- nucleoside
- nucleotides
- polymerase chain reaction (PCR)
- replication
- ribonucleic acid (RNA)
- ribosomal RNA (rRNA)
- Sanger dideoxy method
- sense strand
- small RNAs
- transcription
- transfer RNA (tRNA)
- translation


Key Terms -


Summary -

DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) are biological polymers that act as chemical carriers of an organism's genetic information. Enzyme-catalyzed hydrolysis of nucleic acids yields nucleotides, the monomer units from which RNA and DNA are constructed. Further enzyme-catalyzed hydrolysis of the nucleotides yields nucleosides plus phosphate. Nucleosides, in turn, consist of a purine or pyrimidine base linked to the C1 of an aldopentose sugar-ribose in RNA and 2-deoxyribose in DNA. The nucleotides are joined by phosphate links between the $5^{\prime}$ phosphate of one nucleotide and the $3^{\prime}$ hydroxyl on the sugar of another nucleotide.


Summary -

Molecules of DNA consist of two complementary polynucleotide strands held together by hydrogen bonds between heterocyclic bases on the different strands and coiled into a double helix. Adenine and thymine form hydrogen bonds to each other, as do cytosine and guanine.


Summary -

Three processes take place in deciphering the genetic information of DNA:


Summary -

- Replication of DNA is the process by which identical DNA copies are made. The DNA double helix unwinds, complementary deoxyribonucleotides line up in order, and two new DNA molecules are produced.
- Transcription is the process by which RNA is produced to carry genetic information from the nucleus to the ribosomes. A short segment of the DNA double helix unwinds, and complementary ribonucleotides line up to produce messenger RNA (mRNA).
- Translation is the process by which mRNA directs protein synthesis. Each mRNA is divided into codons, ribonucleotide triplets that are recognized by small amino acid-carrying molecules of transfer RNA (tRNA), which deliver the appropriate amino acids needed for protein synthesis.


Summary -

Sequencing of DNA is carried out by the Sanger dideoxy method, and small DNA segments can be synthesized in the laboratory by automated instruments. Small amounts of DNA can be amplified by factors of $10^{6}$ using the polymerase chain reaction (PCR).


Additional Problems -

Visualizing Chemistry
PROBLEM Identify the following bases, and tell whether each is found in DNA, RNA, or both:
28-13 (a)
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1031.jpg?height=359&width=470&top_left_y=1523&top_left_x=486)
(b)
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1031.jpg?height=359&width=289&top_left_y=1523&top_left_x=1029)
(c)
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1031.jpg?height=377&width=291&top_left_y=1530&top_left_x=1383)


Additional Problems -

PROBLEM Identify the following nucleotide, and tell how it is used:


28-14 -


28-14 -

PROBLEM Amine bases in nucleic acids can react with alkylating agents in typical $\mathrm{S}_{\mathrm{N}} 2$ reactions. Look at the
28-15 following electrostatic potential maps, and tell which is the better nucleophile, guanine or adenine. The reactive positions in each are indicated.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1032.jpg?height=394&width=438&top_left_y=345&top_left_x=426)


28-14 -

9-Methylguanine
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1032.jpg?height=435&width=405&top_left_y=365&top_left_x=941)


Mechanism Problems -

PROBLEM The final step in DNA synthesis is deprotection by treatment with aqueous ammonia. Show the
28-16 mechanisms by which deprotection occurs at the points indicated in the following structure:
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1032.jpg?height=667&width=681&top_left_y=1057&top_left_x=413)


Mechanism Problems -

PROBLEM The final step in the metabolic degradation of uracil is the oxidation of malonic semialdehyde to
28-17 give malonyl CoA. Propose a mechanism.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1032.jpg?height=305&width=668&top_left_y=1864&top_left_x=417)


Mechanism Problems -

PROBLEM One of the steps in the biosynthesis of a nucleotide called inosine monophosphate is the formation 28-18 of aminoimidazole ribonucleotide from formylglycinamidine ribonucleotide. Propose a mechanism.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1033.jpg?height=434&width=776&top_left_y=184&top_left_x=414)


Mechanism Problems -

PROBLEM One of the steps in the metabolic degradation of guanine is hydrolysis to give xanthine. Propose a 28-19 mechanism.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1033.jpg?height=337&width=800&top_left_y=737&top_left_x=413)


Mechanism Problems -

PROBLEM One of the steps in the biosynthesis of uridine monophosphate is the reaction of aspartate with
28-20 carbamoyl phosphate to give carbamoyl aspartate followed by cyclization to form dihydroorotate. Propose mechanisms for both steps.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1033.jpg?height=532&width=1066&top_left_y=1233&top_left_x=416)


General Problems -

PROBLEM Human brain natriuretic peptide (BNP) is a small peptide of 32 amino acids used in the treatment
28-21 of congestive heart failure. How many nitrogen bases are present in the DNA that codes for BNP?
PROBLEM Human and horse insulin both have two polypeptide chains, with one chain containing 21 amino
28-22 acids and the other containing 30 amino acids. They differ in primary structure at two places. At position 9 in one chain, human insulin has Ser and horse insulin has Gly; at position 30 in the other chain, human insulin has Thr and horse insulin has Ala. How must the DNA for the two insulins differ?


General Problems -

PROBLEM The DNA of sea urchins contains about $32 \%$ A. What percentages of the other three bases would you
28-23 expect in sea urchin DNA? Explain.
PROBLEM The codon UAA stops protein synthesis. Why does the sequence UAA in the following stretch of 28-24 mRNA not cause any problems?


General Problems -

PROBLEM Which of the following base sequences would most likely be recognized by a restriction 28-25 endonuclease? Explain.
(a) GAATTC
(b) GATTACA
(c) CTCGAG


General Problems -

PROBLEM For what amino acids do the following ribonucleotide triplets code?
28-26
(a) AAU
(b) GAG
(c) UCC
(d) CAU


General Problems -

PROBLEM From what DNA sequences were each of the mRNA codons in Problem 28-26 transcribed? 28-27

PROBLEM What anticodon sequences of tRNAs are coded for by the codons in Problem 28-26? 28-28


General Problems -

PROBLEM Draw the complete structure of the ribonucleotide codon UAC. For what amino acid does this 28-29 sequence code?


General Problems -

PROBLEM Draw the complete structure of the deoxyribonucleotide sequence from which the mRNA codon in 28-30 Problem 28-29 was transcribed.


General Problems -

PROBLEM Give an mRNA sequence that will code for the synthesis of metenkephalin. 28-31
Tyr-Gly-Gly-Phe-Met


General Problems -

PROBLEM Give an mRNA sequence that will code for the synthesis of angiotensin II. 28-32


General Problems -

> Asp-Arg-Val-Tyr-Ile-His-Pro-Phe


General Problems -

PROBLEM What amino acid sequence is coded for by the following DNA coding strand (sense strand)?
28-33
(5') CTT-CGA-CCA-GAC-AGC-TTT (3')

PROBLEM What amino acid sequence is coded for by the following mRNA base sequence?
28-34
(5') CUA-GAC-CGU-UCC-AAG-UGA (3')
PROBLEM If the DNA coding sequence -CAA-CCG-GAT- were miscopied during replication and became -CGA-28-35 CCG-GAT-, what effect would there be on the sequence of the protein produced?


General Problems -

PROBLEM Show the steps involved in a laboratory synthesis of the DNA fragment with the sequence CTAG. 28-36


General Problems -

PROBLEM Draw the structure of cyclic adenosine monophosphate (cAMP), a messenger involved in the 28-37 regulation of glucose production in the body. Cyclic AMP has a phosphate ring connecting the $3^{\prime}$ and 5 '-hydroxyl groups on adenosine.


General Problems -

PROBLEM Valganciclovir, marketed as Valcyte, is an antiviral agent used for the treatment of cytomegalovirus.
28-38 Called a prodrug, valganciclovir is inactive by itself but is rapidly converted in the intestine by hydrolysis of its ester bond to produce an active drug, called ganciclovir, along with an amino acid.
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General Problems -

Valganciclovir
(a) What amino acid is produced by hydrolysis of the ester bond in valganciclovir?
(b) What is the structure of ganciclovir?
(c) What atoms present in the nucleotide deoxyguanine are missing from ganciclovir?
(d) What role do the atoms missing from deoxyguanine play in DNA replication?
(e) How might valganciclovir interfere with DNA synthesis?


CHAPTER 29 -

The Organic Chemistry of Metabolic Pathways
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1036.jpg?height=1015&width=1064&top_left_y=360&top_left_x=674)


CHAPTER 29 -

FIGURE 29.1 Acyl CoA dehydrogenase is an enzyme that catalyzes the introduction of a $\mathbf{C =} \mathbf{C}$ double bond into fatty acids during their metabolism. (credit: modification of image from the RCSB PDF (rcsb.org) of PDB ID 2WBI. Muniz, J.R.C., Guo, K., Savitsky, P., Roos, A., Yue, W., Pilka, E., Vondelft, F., Edwards, A.M., Bountra, C., Arrowsmith, C.H., Weigelt, J., Oppermann, U. Crystal structure of human Acyl-CoA dehydrogenase 11)


CHAPTER CONTENTS -

29.1 An Overview of Metabolism and Biochemical Energy
29.2 Catabolism of Triacylglycerols: The Fate of Glycerol
29.3 Catabolism of Triacylglycerols: $\beta$-Oxidation
29.4 Biosynthesis of Fatty Acids
29.5 Catabolism of Carbohydrates: Glycolysis
29.6 Conversion of Pyruvate to Acetyl CoA
29.7 The Citric Acid Cycle
29.8 Carbohydrate Biosynthesis: Gluconeogenesis
29.9 Catabolism of Proteins: Deamination
29.10 Some Conclusions about Biological Chemistry


CHAPTER CONTENTS -

WHY THIS CHAPTER? In this chapter, we'll look at some of the pathways by which organisms carry out their chemistry, focusing primarily on how they metabolize fats and carbohydrates. The treatment will be far from complete, but it should give you an idea of the kinds of processes that occur.


CHAPTER CONTENTS -

Anyone who wants to understand or contribute to the revolution now taking place in the biological sciences must first understand life processes at the molecular level. This understanding, in turn, must be based on a detailed knowledge of the chemical reactions and pathways used by living organisms. Just knowing what occurs is not enough; it's also necessary to understand how and why organisms use the chemistry they do.


CHAPTER CONTENTS -

Biochemical reactions are not mysterious. Even though the biological reactions that take place in living organisms often appear complicated, they follow the same rules of reactivity as laboratory reactions and they operate by the same mechanisms.


CHAPTER CONTENTS -

Some of the molecules we'll be encountering are substantially larger and more complex than those we've been dealing with thus far. But don't be intimidated; keep your focus on the parts of the molecules where changes occur, and ignore the parts where nothing changes. The reactions themselves are exactly the same additions, eliminations, substitutions, carbonyl condensations, and so forth, that we've been dealing with all along. By the end of this chapter, it should be clear that the chemistry of living organisms is organic chemistry.


CHAPTER CONTENTS - 29.1 An Overview of Metabolism and Biochemical Energy

The many reactions that occur in the cells of living organisms are collectively called metabolism. The pathways that break down larger molecules into smaller ones are called catabolism, and the pathways that synthesize larger biomolecules from smaller ones are known as anabolism. Catabolic reaction pathways are usually exergonic and release energy, while anabolic pathways are often endergonic and absorb energy. Catabolism can be divided into the four stages shown in FIGURE 29.2.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1037.jpg?height=1432&width=1499&top_left_y=889&top_left_x=313)


FIGURE 29.2 An overview of catabolic pathways for the degradation of food and the production of biochemical energy. The ultimate products of food catabolism are $\mathrm{CO}_{2}$ and $\mathrm{H}_{2} \mathrm{O}$, with the energy released in the citric acid cycle used to drive the endergonic synthesis of adenosine triphosphate (ATP) from adenosine diphosphate (ADP) plus hydrogen phosphate ion, $\mathrm{HOPO}_{3}{ }^{2-}$.


In the first catabolic stage, commonly called digestion, food is broken down in the mouth, stomach, and small intestine by hydrolysis of ester, acetal (glycoside), and amide (peptide) bonds to yield fatty acids, simple sugars,
and amino acids. These smaller molecules are then absorbed and further degraded in the second stage of catabolism to yield acetyl groups attached by a thioester bond to the large carrier molecule, coenzyme A. The resultant compound, acetyl coenzyme A (acetyl CoA), is a key substance in the metabolism of food molecules and in many other biological pathways. As noted in Section 21.8, the acetyl group in acetyl CoA is linked to the sulfur atom of phosphopantetheine, which is itself linked to adenosine 3',5'-bisphosphate.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1038.jpg?height=448&width=1049&top_left_y=432&top_left_x=538)


Acetyl CoA-a thioester -

Acetyl groups are oxidized inside cellular mitochondria in the third stage of catabolism, the citric acid cycle, to yield $\mathrm{CO}_{2}$. (We'll see the details of the process in Section 29.7.) Like most oxidations, this stage releases a large amount of energy, which is used in the fourth stage, the electron-transport chain, to accomplish the endergonic phosphorylation of adenosine diphosphate (ADP) with hydrogen phosphate ion ( $\mathrm{HOPO}_{3}{ }^{2-}$, abbreviated $\mathrm{P}_{\mathrm{i}}$ ) to give adenosine triphosphate (ATP).


Acetyl CoA-a thioester -

As the final result of food catabolism, ATP has been called the "energy currency" of the cell. Catabolic reactions "buy" ATP with the energy they release to synthesize it from ADP and hydrogen phosphate ion. Anabolic reactions then spend the ATP by transferring a phosphate group to another molecule, thereby regenerating ADP. Energy production and use in living organisms thus revolves around the ATP $\rightleftarrows$ ADP interconversion.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1038.jpg?height=508&width=1611&top_left_y=1340&top_left_x=271)
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breaking a $\mathrm{C}-\mathrm{O}$ bond and forming a carboxylic ester, ROCOR' (Section 21.5), phosphoric acid anhydrides react with alcohols by breaking a $\mathrm{P}-\mathrm{O}$ bond and forming a phosphate ester, $\mathrm{ROPO}_{3}{ }^{2-}$. The reaction is, in effect, a nucleophilic acyl substitution at phosphorus. Note that phosphorylation reactions with ATP generally require the presence of a divalent metal cation in the enzyme, usually $\mathrm{Mg}^{2+}$, to form a Lewis acid-base complex with the phosphate oxygen atoms and to neutralize negative charge.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1039.jpg?height=700&width=1313&top_left_y=192&top_left_x=406)


Acetyl CoA-a thioester -

How does the body use ATP? Recall from Section 6.7 that the free-energy change $\Delta G$ must be negative and energy must be released for a reaction to be energetically favorable and occur spontaneously. If $\Delta G$ is positive, the reaction is energetically unfavorable and the process can't occur spontaneously.


Acetyl CoA-a thioester -

For an energetically unfavorable reaction to occur, it must be "coupled" to an energetically favorable reaction so that the overall free-energy change for the two reactions together is favorable. To understand what it means for reactions to be coupled, imagine that reaction 1 does not occur to any reasonable extent because it has a small equilibrium constant and is energetically unfavorable; that is, the reaction has $\Delta G>0$.


Acetyl CoA-a thioester -

$$
(1) \mathbf{A}+m \longleftrightarrow \mathbf{B}+n \quad \Delta G>0
$$


Acetyl CoA-a thioester -

where $\mathbf{A}$ and $\mathbf{B}$ are the biochemically "important" substances while $m$ and $n$ are enzyme cofactors, $\mathrm{H}_{2} \mathrm{O}$, or other small molecules.


Acetyl CoA-a thioester -

Imagine also that product $n$ can react with substance $o$ to yield $p$ and $q$ in a second, highly favorable reaction that has a large equilibrium constant and $\Delta G \ll 0$.
(2) $n+o \rightleftarrows p+q \quad \Delta G \ll 0$


Acetyl CoA-a thioester -

Taking the two reactions together, they share, or are coupled through, the common intermediate $n$, which is a product in the first reaction and a reactant in the second. When even a tiny amount of $n$ is formed in reaction 1 , it undergoes essentially complete conversion in reaction 2 , thereby removing it from the first equilibrium and forcing reaction 1 to continually replenish $n$ until reactant $\mathbf{A}$ is gone. That is, the two reactions added together have a favorable $\Delta G<0$, and we say that the favorable reaction 2 "drives" the unfavorable reaction 1 . Because the two reactions are coupled through $n$, the transformation of $\mathbf{A}$ to $\mathbf{B}$ becomes favorable.


Acetyl CoA-a thioester -

| (1) $\mathbf{A}+m \longleftrightarrow \mathbf{B}+\not h$ | $\Delta G>0$ |
| :--- | :--- |
| (2) $\not \propto+o \rightleftarrows p+q$ | $\Delta G \ll 0$ |
| Net: $\mathbf{A}+m+o \rightleftarrows \mathbf{B}+p+q$ | $\Delta G<0$ |


Acetyl CoA-a thioester -

For an example of two reactions that are coupled, look at the phosphorylation reaction of glucose to yield glucose 6-phosphate plus water, an important step in the breakdown of dietary carbohydrates.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1039.jpg?height=163&width=1291&top_left_y=2133&top_left_x=414)


Acetyl CoA-a thioester -

Glucose
Glucose 6-phosphate
The reaction of glucose with $\mathrm{HOPO}_{3}{ }^{2-}$ does not occur spontaneously because it is energetically unfavorable, with $\Delta G^{\circ{ }^{\prime}}=13.8 \mathrm{~kJ} / \mathrm{mol}$. (The standard free-energy change for a biological reaction is denoted $\Delta G^{\circ}$ and refers to a process in which reactants and products have a concentration of 1.0 M in a solution with $\mathrm{pH}=7$.) At the same time, however, the reaction of water with ATP to yield ADP plus $\mathrm{HOPO}_{3}{ }^{2-}$ is strongly favorable, with $\Delta G^{\circ}$
$=-30.5 \mathrm{~kJ} / \mathrm{mol}$. When the two reactions are coupled, glucose reacts with ATP to yield glucose 6-phosphate plus ADP in a reaction that is favorable by about $16.7 \mathrm{~kJ} / \mathrm{mol}(4.0 \mathrm{kcal} / \mathrm{mol})$. That is, ATP drives the phosphorylation reaction of glucose.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1040.jpg?height=185&width=1256&top_left_y=355&top_left_x=429)


Acetyl CoA-a thioester -

It's this ability to drive otherwise unfavorable phosphorylation reactions that makes ATP so useful. The resultant phosphates are much more reactive as leaving groups in nucleophilic substitutions and eliminations than the alcohols they're derived from and are therefore more chemically useful.


Acetyl CoA-a thioester -

PROBLEM One of the steps in fat metabolism is the reaction of glycerol (1,2,3-propanetriol) with ATP to yield 29-1 glycerol 1-phosphate. Write the reaction, and draw the structure of glycerol 1-phosphate.


Acetyl CoA-a thioester - 29.2 Catabolism of Triacylglycerols: The Fate of Glycerol

The metabolic breakdown of triacylglycerols begins with their hydrolysis in the stomach and small intestine to yield glycerol plus fatty acids. The reaction is catalyzed by a lipase, whose mechanism is shown in FIGURE 29.3. The active site of the enzyme contains a catalytic triad of aspartic acid, histidine, and serine residues, which act cooperatively to provide the necessary acid and base catalysis for the individual steps. Hydrolysis is accomplished by two sequential nucleophilic acyl substitution reactions, one that covalently binds an acyl group to the side chain -OH of a serine residue on the enzyme and a second that frees the fatty acid from the enzyme.


FIGURE 29.3 MECHANISM -

Mechanism for the action of lipase. The active site of the enzyme contains a catalytic triad of aspartic acid, histidine, and serine, which react cooperatively to carry out two nucleophilic acyl substitution reactions. Individual steps are explained in the text.
(1) The enzyme active site contains an aspartic acid, a histidine, and a serine. First, histidine acts as a base to deprotonate the -OH group of serine, with the negatively charged carboxylate of aspartic acid stabilizing the nearby histidine cation that results. Serine then adds to the carbonyl group of the triacylglycerol, yielding a tetrahedral intermediate.


FIGURE 29.3 MECHANISM -

This intermediate expels a diacylglycerol as leaving group in a nucleophilic acyl substitution reaction, giving an acyl enzyme. The diacylglycerol is protonated by the histidine cation.


FIGURE 29.3 MECHANISM -

3 Histidine deprotonates a water molecule, which adds to the acyl group. A tetrahedral intermediate is again formed, and the histidine cation is again stabilized by the nearby carboxylate.
4) The tetrahedral intermediate expels the serine as leaving group in a second nucleophilic acyl substitution reaction, yielding a free fatty acid. The serine accepts a proton from histidine, and the enzyme has now returned to its starting structure.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1041.jpg?height=660&width=722&top_left_y=451&top_left_x=1032)


FIGURE 29.3 MECHANISM -

Tetrahedral intermediate
(2)
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1041.jpg?height=351&width=646&top_left_y=1245&top_left_x=1051)
(3) $\downarrow \mathrm{H}_{2} \mathrm{O}$
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1041.jpg?height=226&width=603&top_left_y=1703&top_left_x=1051)

Tetrahedral intermediate


(4) -


(4) -

Enzyme
Fatty acid


(4) -

Steps 1-2 of FIGURE 29.3: Acyl Enzyme Formation
The first nucleophilic acyl substitution step-reaction of the triacylglycerol with the active-site serine to give an acyl enzyme-begins with deprotonation of the serine alcohol by histidine to form the more strongly nucleophilic alkoxide ion. This proton transfer is facilitated by a nearby side-chain carboxylate anion of aspartic acid, which makes the histidine more basic and stabilizes the resultant histidine cation by electrostatic interactions. The deprotonated serine adds to a carbonyl group of a triacylglycerol to give a tetrahedral intermediate, which expels a diacylglycerol as a leaving group and produces an acyl enzyme. This step is catalyzed by a proton transfer from histidine to make the leaving group a neutral alcohol.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1042.jpg?height=698&width=1397&top_left_y=562&top_left_x=364)

Steps 3-4 of FIGURE 29.3: Hydrolysis
The second nucleophilic acyl substitution step hydrolyzes the acyl enzyme and gives the free fatty acid by a mechanism analogous to that of the first two steps. Water is deprotonated by histidine to give hydroxide ion, which adds to the enzyme-bound acyl group. The tetrahedral intermediate then expels the neutral serine residue as the leaving group, freeing the fatty acid and returning the enzyme to its active form.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1042.jpg?height=712&width=1489&top_left_y=1525&top_left_x=313)


(4) -

The fatty acids released on triacylglycerol hydrolysis are transported to mitochondria and degraded to acetyl CoA, while the glycerol is carried to the liver for further metabolism. In the liver, glycerol is first phosphorylated by reaction with ATP and then oxidized by $\mathrm{NAD}^{+}$. The resulting dihydroxyacetone phosphate (DHAP) enters the carbohydrate glycolysis pathway, which we'll discuss in Section 29.5.
![](https://cdn.mathpix.com/cropped/2025_02_06_5556f7656cbdcaa0f9a7g-1043.jpg?height=435&width=1220&top_left_y=192&top_left_x=447)


(4) -

You might note that C2 of glycerol is a prochiral center (Section 5.11) with two identical "arms." As is typical for enzyme-catalyzed reactions, the phosphorylation of glycerol is selective. Only the pro- $R$ arm undergoes reaction, although this can't be predicted in advance.


(4) -

Note also that the phosphorylation product is named $s n$-glycerol 3-phosphate, where the $s n$ - prefix means "stereospecific numbering." In this convention, the molecule is drawn as a Fischer projection with the -OH group at C2 pointing to the left and the glycerol carbon atoms numbered from the top.


(4) - 29.3 Catabolism of Triacylglycerols: $\boldsymbol{\beta}$-Oxidation

The fatty acids that result from triacylglycerol hydrolysis are converted into thioesters with coenzyme A and then catabolized by a repetitive four-step sequence of reactions called the $\boldsymbol{\beta}$-oxidation pathway, shown in FIGURE 29.4. Each passage along the pathway results in the cleavage of an acetyl group from the end of the fatty-acid chain, until the entire molecule is ultimately degraded. As each acetyl group is produced, it enters the citric acid cycle and is further degraded to $\mathrm{CO}_{2}$, as we'll see in Section 29.7.