title: Laminin alterations after in vitro nonenzymatic glycosylation.
abstract: Laminin, a basement membrane protein derived from the matrix of the Engelbreth-Holm-Swarm murine tumor, was nonenzymatically glycosylated in vitro in the presence of increasing glucose concentrations. The amount of glucose incorporated per laminin molecule was shown to be proportional to the molarity of glucose used. Nonenzymatic glycosylation resulted in formation of cross-links and alterations of the cruciform shape of laminin molecules; these alterations were dramatic when high concentrations of glucose were used. One of the functions of laminin, the process of self-assembly, was shown to be impaired after in vitro nonenzymatic glycosylation. Glucose incorporation resulted in a dramatic decrease of long-to-long laminin dimers, which normally form during the initial steps of assembly. Furthermore, nonenzymatic glycosylation of laminin reduced its ability to self-associate into complexes larger than dimers, as judged by turbidimetry. The observed decrease of maximal turbidity was proportional to the degree of nonenzymatic glycosylation. Aminoguanidine, which has been suggested to inhibit cross-link formation, was shown to restore to a large extent the shape of laminin, the percentage of long-to-long arm dimers, and the maximal turbidity when included in the mixtures of laminin and glucose. These data suggest that structural and functional alterations of laminin may be primarily due to formation of cross-links. Such modifications of laminin (along with our basement membrane components) may contribute to the morphological and physiological changes observed in basement membranes under diabetic conditions.
authors.affiliation: Department of Laboratory, University of Minnesota Medical School, Minneapolis.
authors.forename: A S
authors.initials: AS
authors.lastname: Charonis
authors.forename: L A
authors.initials: LA
authors.lastname: Reger
authors.forename: J E
authors.initials: JE
authors.lastname: Dege
authors.forename: K
authors.initials: K
authors.lastname: Kouzi-Koliakos
authors.forename: L T
authors.initials: LT
authors.lastname: Furcht
authors.forename: R M
authors.initials: RM
authors.lastname: Wohlhueter
authors.forename: E C
authors.initials: EC
authors.lastname: Tsilibary
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D006146:Guanidines / Q000494:pharmacology; D007700:Kinetics; D007797:Laminin / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D008239:Lysine; D051379:Mice; D009374:Neoplasms, Experimental / Q000378:metabolism; D009391:Nephelometry and Turbidimetry
chemical_list: D006146:Guanidines; D007797:Laminin; D005947:Glucose; D008239:Lysine; C004479:pimagedine