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  • Mouse fibroblasts null for the long isoform of β1,4-galactosyltransferase-I show defective cell-matrix interactions.

Mouse fibroblasts null for the long isoform of β1,4-galactosyltransferase-I show defective cell-matrix interactions.

Biochemical and biophysical research communications (2016-08-25)
Brooke H Elder, Barry D Shur
ABSTRACT

β1,4 Galactosyltransferase-I (GalT-I) is expressed as two nearly identical polypeptides that differ only in the length of their cytoplasmic domains. The longer isoform has been implicated as a cell surface receptor for extracellular glycoside ligands, such as laminin. To more stringently test the function of the long GalT-I isoform during cell interactions with laminin, we created multiple independent fibroblastic cell lines that fail to express the long isoform, but which express the short GalT-I isoform normally and appear to have normal intracellular galactosylation. Cells devoid of the long GalT-I isoform are unable to adhere and spread on laminin substrates as well as control cells, but retain near normal interactions with fibronectin, which do not rely upon surface GalT-I function. The loss of the long GalT-I isoform also leads to a loss of actin stress fibers, focal adhesions and rac GTPase activation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Neuraminidase from Clostridium perfringens (C. welchii), Type VIII, lyophilized powder, 10-20 units/mg protein (using 4MU-NANA), 3.5-8.0 units/mg protein (mucin)
Sigma-Aldrich
Anti-Rac1 Antibody, clone 23A8, clone 23A8, Upstate®, from mouse
Sigma-Aldrich
Anti-Integrin α6 Antibody, clone NKI-GoH3, clone NKI-GoH3, Chemicon®, from rat