title: High capacity, low affinity Ca2+ binding of chromogranin A. Relationship between the pH-induced conformational change and Ca2+ binding property.
abstract: Chromogranin A, the major intravesicular protein of adrenal chromaffin granules, bound Ca2+ in a pH-dependent manner. Both the maximal binding and affinity of chromogranin A for Ca2+ were dependent on pH. Chromogranin A bound 670 nmol of Ca2+/mg (32 mol/mol) and 1150 nmol of Ca2+/mg (55 mol/mol) at pH 7.5 and 5.5, respectively, with dissociation constants (Kd) of 2.7 and 4 mM. This pH dependence probably reflects different conformations of the protein at the two pH values. Conformational differences of chromogranin A at two different pH values were demonstrated by limited tryptic digestion patterns confirming previous results obtained by circular dichroism spectroscopy (Yoo, S. H., and Albanesi, J. P. (1990) J. Biol. Chem. 265, 14414-14421). Sedimentation equilibrium studies revealed the native molecular mass of chromogranin A to be 100 kDa at pH 7.5 and 192 kDa at pH 5.5, indicating dimeric and tetrameric states of the protein at the two pH levels. We postulate that the pH- and Ca2(+)-induced conformational changes of chromogranin A may have a role both in the regulation of Ca2+ release of chromaffin granules and in the early stages of secretory vesicle biogenesis.
authors.affiliation: University of Texas Southwestern Medical Center, Dallas 75235.
authors.forename: S H
authors.initials: SH
authors.lastname: Yoo
authors.forename: J P
authors.initials: JP
authors.lastname: Albanesi
mesh_terms: D000311:Adrenal Glands / Q000737:chemistry; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002850:Chromatography, Gel; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D014357:Trypsin
chemical_list: C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D014357:Trypsin; D002118:Calcium