Title

Intrinsic tryptophan fluorescence measurements suggest that polylactosaminyl glycosylation affects the protein conformation of the gelatin‐binding domain from human placental fibronectin

Document Type

Article

Publication Date

1-1-1990

Abstract

Glycosylation can affect the physical and biochemical properties of the polypeptide chain in glycoproteins. Asparagine‐N‐linked polylactosaminyl glycosylation of the chymotryptic 44‐kDa gelatin‐binding domain from human placental fibronectin confers protease resistance [Zhu, B. C. R., Fisher, S. F., Panda, H., Calaycay, J., Shively, J. E. & Laine, R. A. (1984) J. Biol. Chem. 259, 3962–3970] and weakens the binding to gelatin [Zhu, B. C. R. & Laine, R. A. (1985) J. Biol. Chem. 260, 4041–4045]. Intrinsic tryptophan fluorescence of the gelatinbinding domain was used to probe glycosylation‐dependent protein conformation changes. In gelatin‐binding fragments containing incrementally smaller polylactosamine oligosaccharides, the fluorescence intensity progressively decreased and the emission spectrum shifted about 7 nm to the blue. Removal of the polylactosamine chains from a highly glycosylated fragment with endo‐β‐galactosidase from Escherichia freundii also quenched the protein fluorescence. The fluorescence lifetimes did not appear to be affected by the extent of glycosylation, suggesting static quenching of the tryptophan emission in the low glycosylated fragments. Acrylamide quenching studies showed that the accessibility of the tryptophans to small solutes was not altered by glycosylation. The steadystate emission anisotropy increased with decreasing polylactosamine chain length. The results indicate that the polylactosamine chains alter the tryptophan environments in the gelatin‐binding domain, probably by changing the polypeptide conformation. These putative protein conformation changes may be partially responsible for the altered gelatin binding, protease resistance, and cell adhesion functions of fetal tissue fibronectin. Copyright © 1990, Wiley Blackwell. All rights reserved

Publication Source (Journal or Book title)

European Journal of Biochemistry

First Page

509

Last Page

516

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