Abstract
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.
Highlights
Identification of the Uridine 5’-Diphosphoglucose (UDP-Glc) Binding Subunit of Cellulose Synthase in Acetobacter xylinum Using the Photoaffinity Probe 5-Azido-UDP-Glc*
The reversible and photocatalyzed binding of this photoprobe 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
There are three major requirements for demonstrating the validity of specific protein binding in photolabeling studies: (a) only a specific protein(s) in an enzyme preparation is photolabeled with the photoprobe; (b) specific photoincorpor
Summary
Vol 265, No 9, Issue of March 25, pp. 4782-4784, 1990 0 1990 by The American Society for Biochemistry and Molecular Biology, Inc. 4782-4784, 1990 0 1990 by The American Society for Biochemistry and Molecular Biology, Inc. Identification of the Uridine 5’-Diphosphoglucose (UDP-Glc) Binding Subunit of Cellulose Synthase in Acetobacter xylinum Using the Photoaffinity Probe 5-Azido-UDP-Glc*. Glc) has been used to identify the UDP-Glc binding subunit of the cellulose synthase from Acetobucter xylinum strain ATCC 53582. The reversible and photocatalyzed binding of this photoprobe 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. Cyclic diguanylic acid, N-Gx, a crude preparation of cellulose synthase activator. In this report, [/?-32P]5-N3UDP-Glc was used to and effectively photolabel an 83-kDa polypeptide in a purified cellulose synthase preparation from A. xylinum strain ATCC
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