title: The R-state proinsulin and insulin hexamers mimic the carbonic anhydrase active site.
abstract: The cobalt(II)-substituted proinsulin and insulin hexamers have been studied in solution via electronic absorption spectroscopy. Hexameric proinsulin is shown to undergo the phenol-induced T6 to R6 conformational transition in a manner analogous to that previously established for insulin. In the absence of coordinating anions, the coordination spheres of the Co(II) ions in the proinsulin and insulin R6 hexamers comprise identical pseudotetrahedral arrangements of 3 histidine residues and 1 hydroxide ion. At alkaline pH, the visible absorption spectrum of the phenol-induced R6 Co(II) center is strikingly similar to the distinctive spectrum of the alkaline form of Co(II)-carbonic anhydrase. Exogenous ligands may coordinate to the Co(II) ions of the R6 proinsulin and insulin hexamers via replacement of the hydroxide ion, forming pseudotetrahedral adducts possessing characteristic spectra. The binding affinity of such ligands is shown to be strongly pH-dependent. The data presented establish that, although the Co(II)-substituted proinsulin and insulin R6 hexamers lack enzyme-like activity, these species duplicate spectrochemical characteristics of the Co(II)-carbonic anhydrase active site that are believed to be important signatures of carbonic anhydrase catalytic function.
authors.affiliation: Department of Biochemistry, University of California, Riverside 92521.
authors.forename: M L
authors.initials: ML
authors.lastname: Brader
authors.forename: N C
authors.initials: NC
authors.lastname: Kaarsholm
authors.forename: M F
authors.initials: MF
authors.lastname: Dunn
mesh_terms: D000818:Animals; D001665:Binding Sites; D002256:Carbonic Anhydrases / Q000378:metabolism*; D003035:Cobalt / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D046911:Macromolecular Substances; D008958:Models, Molecular; D011384:Proinsulin / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013053:Spectrophotometry; D013552:Swine
chemical_list: D007328:Insulin; D046911:Macromolecular Substances; D003035:Cobalt; D011384:Proinsulin; D002256:Carbonic Anhydrases