Identification of the uridine 5'-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photoaffinity probe 5-azido-UDP-Glc.
Photoaffinity labeling of purified cellulose synthase with [beta-32P]5-azidouridine 5'-diphosphoglucose (UDP-Glc) has been used to identify the UDP-Glc binding subunit of the cellulose synthase from Acetobacter xylinum strain ATCC 53582. The results showed exclusive labeling of an 83-kDa polypeptide. Photoinsertion of [beta-32P]5-azido-UDP-Glc is stimulated by the cellulose synthase activator, bis-(3'----5') cyclic diguanylic acid. Addition of increasing amounts of UDP-Glc prevents photolabeling of the 83-kDa polypeptide. The reversible and photocatalyzed binding of this photoprobe also showed saturation kinetics. These studies demonstrate that the 83-kDa polypeptide is the catalytic subunit of the cellulose synthase in A. xylinum strain ATCC 53582.
Lin
F C
FC
Department of Botany, University of Texas, Austin 78713-7640.
Brown
R M
RM
Drake
R R
RR
Haley
B E
BE
D000345:Affinity Labels / Q000378:metabolism*; D029681:Arabidopsis Proteins; D001345:Autoradiography; D001386:Azides / Q000378:metabolism*; D001665:Binding Sites; D002462:Cell Membrane / Q000201:enzymology; D041822:Gluconacetobacter xylinus / Q000201:enzymology*; D005964:Glucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010761:Phosphorus Radioisotopes; D014532:Uridine Diphosphate Glucose / Q000031:analogs & derivatives / Q000378:metabolism*; D014539:Uridine Diphosphate Sugars / Q000378:metabolism*
D000345:Affinity Labels; D029681:Arabidopsis Proteins; D001386:Azides; D046911:Macromolecular Substances; D010761:Phosphorus Radioisotopes; D014539:Uridine Diphosphate Sugars; C060227:5-azidouridine 5'-diphosphoglucose; D005964:Glucosyltransferases; C457769:PRC1 protein, Arabidopsis; C028634:cellulose synthase (UDP-forming); D014532:Uridine Diphosphate Glucose