Site-directed mutagenesis study of the role of histidine residues in the neutral-to-basic transition of human serum albumin.
Site-directed mutagenesis was used to study the role of histidine residues located in domain I in the neutral-to-basic (N-B) transition of human serum albumin (HSA). Based on a previous study of the N-B transition by means of proton NMR, the following recombinant HSA species were synthesized in the yeast species, Pichia pastoris: H9F, H9S, H39F, H39S, H67F, H67S, H105F, H105S, H128F, H128S, H146F, H146S, and wild type HSA. By monitoring the fluorescent intensity of warfarin bound to the above recombinant human serum albumin species as a function of pH, the mutational effect of individual histidine residues on the N-B transition was examined. While H9, H67, H105, H128 and H146 contribute to the transition significantly, H39 appears to have virtually no contribution to the transition. Based on the X-ray crystallographic structure, it is suggested that electrostatic interactions are the principal factor in determining the histidine pK shifts.
Yang
Jinsheng
J
Department of Anatomy, Biochemistry, Physiology and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, 1960 East-West Road., Honolulu, HI 96822, USA.
Ha
Chung-Eun
CE
Bhagavan
Nadhipuram V
NV
D006639:Histidine / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010843:Pichia / Q000737:chemistry / Q000235:genetics; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D012709:Serum Albumin / Q000032:analysis / Q000737:chemistry* / Q000235:genetics
D011994:Recombinant Proteins; D012709:Serum Albumin; D006639:Histidine