Synthetic metal-binding protein surface domains for metal ion-dependent interaction chromatography. II. Immobilization of synthetic metal-binding peptides from metal ion transport proteins as model bioactive protein surface domains.
This preliminary investigation tests the premise that biologically relevant (1) peptide-metal ion interactions, and (2) metal ion-dependent macromolecular recognition events (e.g., peptide-peptide interactions) may be modeled by biomimetic affinity chromatography. Divinylsulfone-activated agarose (6%) was used to immobilize three different synthetic peptides representing metal-binding protein surface domains from the human plasma metal transport protein histidine-rich glycoprotein (HRG). The synthetic peptides represented 1-3 multiple repeat units of the 5-residue sequence (Gly-His-His-Pro-His) found in the C-terminal of HRG. By frontal analyses, immobilized HRG peptides of the type (GHHPH)nG, where n = 1-3, were each found to have a similar binding capacity for both Cu(II) ions and Zn(II) ions (31-38 mumol/ml gel). The metal ion-dependent interaction of a variety of model peptides with each of the immobilized HRG peptide affinity columns demonstrated differences in selectivity despite the similar internal sequence homology and metal ion binding capacity. The immobilized 11-residue HRG peptide was loaded with Cu(II) ions and used to demonstrate selective adsorption and isolation of proteins from human plasma. These results suggest that immobilized metal-binding peptides selected from known solvent-exposed protein surface metal-binding domains may be useful model systems to evaluate the specificity of biologically relevant metal ion-dependent interaction and transfer events in vitro.
Hutchens
T W
TW
US Department of Agriculture/Agricultural Research Service, Houston, TX.
Yip
T T
TT
D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000378:metabolism*; D002413:Cations, Divalent; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008956:Models, Chemical; D008969:Molecular Sequence Data; D011506:Proteins / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*; D000099053:Histidine-Rich Glycoprotein
D001798:Blood Proteins; D002413:Cations, Divalent; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein; D003300:Copper; D015032:Zinc