title: Chemical properties of the functional groups of insulin.
abstract: The method of competitive binding [Kaplan, Stevenson & Hartley (1971) Biochem. J. 124, 289-299] with 1-fluoro-2,4-dinitrobenzene as the labelling reagent [Duggleby & Kaplan (1975) Biochemistry 14, 5168-5175] was used to determine the chemical properties, namely pK and reactivity, of the amino groups, the histidine residues and the tyrosine residues of the dimeric form of pig zinc-free insulin at 20.0 degrees C. The N-terminal glycine residue of the A-chain has a pK of 7.7 and a slightly higher than normal reactivity. The N-terminal phenylalanine residue of the B-chain has a pK of 6.9 and is approximately an order of magnitude more reactive than a corresponding amino group with the same pK value. The lysine epsilon-amino group has an unusually low pK of 7.0 but has approximately the expected reactivity of such a group. In the case of the two histidine and four tyrosine residues only the average properties of each class were determined. The histidine residues have a pK value of approx. 6.6, but, however, their reactivity is at least an order of magnitude greater than that of a free imidazole group. The tyrosine residues have a pK value of approx. 10, but their average reactivities are substantially less than for a free phenolic group. At alkaline pH values above 8 the reactivity of all the functional groups show sharp discontinuities, indicating that insulin is undergoing a structural change that alters the properties of these groups.
mesh_terms: D000089:Acetic Anhydrides / Q000378:metabolism; D000345:Affinity Labels; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D004139:Dinitrofluorobenzene / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism; D013552:Swine
chemical_list: D000089:Acetic Anhydrides; D000345:Affinity Labels; D000596:Amino Acids; D007093:Imidazoles; D007328:Insulin; D007773:Lactates; D004139:Dinitrofluorobenzene