Binding characteristics of a fluorescent analogue of diphosphoglyceric acid to human, amphibian and fish hemoglobins

An attempt was made to establish the binding of N-(2,4-diphosphobenzyl)-1-amino-5-naphthalenesulfonic acid, DIPANS, as an estimator of conformation in the carbonmonoxy (CO)-hemoglobins (Hbs) of several vertebrates. DIPANS failed to bind menhaden I, trout I or tuna Hbs which are ligand insensitive. Below a pH of 7.0, DIPANS bound menhaden II, Bufo, Xenopus, and human Hbs with a binding stoichiometry greater than one. The charge of the DIPANS molecule does not control its binding to these Hbs. The binding to human CO-Hb can not be due to Hb conformation. For Xenopus Hbs and menhaden II, conformation predominates DIPANS binding. The binding to CO-Hb of DIPANS, can not be unambiguously attributed to the Hb's quaternary conformation.