Lysyl oxidase-catalyzed cross-linking and insolubilization reactions of Lys-containing polypeptides and synthetic adhesive proteins.
The lysyl oxidase- (LO-) mediated insolubilization reactions of the Lys-containing polypeptides have been examined using poly(L-Lys) with degrees of polymerization (Dps) ranging 1 from 2300, copoly(LysxAlay) (x:y = 1:4, 1:3; 1:1, 2:1, and 3:1), copoly(LysxGlyy) (x:y = 1:1 and 2:1), and synthetic adhesive proteins with sequential repetitive units enriched in the Lys residues, poly(Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Lys), poly(Ala-Gly-Tyr-Gly-Gly-Ala-Lys), and poly(Gly-Gly-Gly-Tyr-Gly-Gly-Tyr-Gly-Lys). All of the substrates were insolubilized by the LO-catalyzed oxidation of the epsilon-amino group in the Lys residues. The Dps of the polypeptide substrates did not affect the kinetic constants, the Km and Vmax values. The Km and Vmax values and the insolubilization rates varied depending on the Lys contents in the substrate polypeptides, which were enriched in Gly and Ala residues. As the Lys content increased, the Km and Vmax values became lower and higher, respectively. The insolubilization rates decreased with increase of the Lys content. The time-dependent changes in the LO-catalyzed aldehyde production, the insolubilization, and remaining LO activity demonstrated that the cross-linking and the insolubilization steps occurred along with LO deactivation, indicating that the enzymatic and chemical processes in the LO-mediated insolubilization occur in order.
Ohkawa
K
K
Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University, Ueda 386-8567, Japan.
Fujii
K
K
Nishida
A
A
Yamauchi
T
T
Ishibashi
H
H
Yamamoto
H
H
D000269:Adhesives / Q000138:chemical synthesis / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D001013:Aorta, Thoracic / Q000201:enzymology; D049872:Bivalvia / Q000201:enzymology; D002384:Catalysis; D002417:Cattle; D003432:Cross-Linking Reagents; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D046911:Macromolecular Substances; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D008249:Protein-Lysine 6-Oxidase / Q000378:metabolism*; D012995:Solubility; D013379:Substrate Specificity
D000269:Adhesives; D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D010455:Peptides; D008249:Protein-Lysine 6-Oxidase; D008239:Lysine