4,4'-Bis (1-anilinonaphthalene 8-sulfonate) (bis-ANS): a new probe of the active site of myosin.

The interaction of myosin subfragment-1 (S-1) with 4,4'-bis(1-anilinonaphthalene 8-sulfonate) (bis-ANS) has been studied by monitoring the fluorescence of the latter when the two components form a complex. Because ATP and ATP analogs partially displace complexed bis-ANS it has also been possible to study interactions of S-1 and nucleotides by using the displacement effect. Approximate values of the parameters of these various interactions have been measured. Some possible applications of bis-ANS have been explored. For example, it provides a very convenient method for obtaining the Michaelis constant, Km, in steady-state S-1 nucleoside triphosphatase; this particular application has also provided some evidence for inferring that in Ca2+ (but not in Mg2+) adenosinetriphosphatase (ATP phosphohydrolase, EC 3.6.1.3) S-1 behaves like a mixture of two components, each with its own Km. Clear energy transfer occurs between tryptophan residues and bound bis-ANS. The fluorescence also suggests that S-1 undergoes some slow relaxations following substrate binding.