Conjugating TNP to ATP renders this nucleotide triphosphate fluorescent and colored whilst allowing it to retain its biological activity. TNP-ATP is thus a  fluorescent analog of ATP. This conjugation is very useful in providing information about interactions between ATP and an ATP-binding protein because TNP-ATP interacts with proteins and enzymes as a substitute for its parent nucleotide, and has a strong binding affinity for most systems that require ATP.
TNP is excited at a wavelength of 408 and 470 nm, and fluoresces in the 530–560 nm range. This is a very useful range of excitation because it is far from where proteins or nucleotides absorb. When TNP-ATP is in water or other aqueous solutions, this emission is very weak. However, once TNP-ATP binds to a protein, there is a dramatic increase in fluorescent intensity. This property enables researchers to study various proteins’ binding interaction with ATP. Thus, with enhanced fluorescence, it can be seen whether a protein binds to ATP.
When TNP-ATP in water is excited at 410 nm, TNP-ATP shows a single fluorescence maximum at 561 nm. This maximum shifts as the fluid's viscosity changes. For example, in N,N-dimethylformamide, instead of having its maxima at 561 nm as in water, the maxima is instead at 533 nm.
Binding to a protein will also change the wavelength of maximal emission, as well as a change in fluorescent intensity. For example, binding to the chemotaxis protein CheA indicates a severalfold enhancement of fluorescence intensity and a blue-shift in wavelength of the maximal emission.
Using this TNP nucleotide analog has been shown in many instances to be superior to traditional radionucleotide-labelling based techniques. The health concerns and the cost associated with the use of radioactive isotopes makes TNP-ATP an attractive alternative.
The first fluorescent ribose-modified ATP is 2’,3’-O-(2,4,7-trinitrocyclohexadienylidene) adenosine 5’triphosphate (TNP-ATP), and was introduced in 1973 by Hiratsuka and Uchida.  TNP-ATP was originally synthesized to investigate the ATP binding site of myosin ATPase. Reports of TNP-ATP’s success in the investigation of this motor protein extended TNP-ATP’s use to other proteins and enzymes. TNP-ATP has now been used as a spectroscopic probe for numerous proteins suspected to have ATP interactions. These include several protein kinases, ATPases, myosin, and other nucleotide binding proteins. Over the past twenty years, there have been hundreds of papers describing TNP-ATP’s use and applications. Many applications involving this fluorescently labeled nucleotide have helped to clarify structure-function relationships of many ATP-requiring proteins and enzymes. There have also been a growing number of papers that display TNP-ATP use as a means of assessing the ATP-binding capacity of various mutant proteins.