Identification of the uridine 5'-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photoaffinity probe 5-azido-UDP-Glc

• Published in: The Journal of biological chemistry Vol. 265; no. 9; pp. 4782 - 4784
• Main Authors: Lin, F.C, Brown, R.M. Jr, Drake, R.R. Jr, Haley, B.E
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 25.03.1990
• Subjects: 550201 - Biochemistry- Tracer Techniques; Acetobacter; AFFINITY; Affinity Labels - metabolism; analogs; Analytical, structural and metabolic biochemistry; Arabidopsis Proteins; AUTORADIOGRAPHY; Azides - metabolism; BACTERIA; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; Binding Sites; Biological and medical sciences; CELL CONSTITUENTS; Cell Membrane - enzymology; CELL MEMBRANES; cellulose; cellulose synthase (UDP-forming); DAYS LIVING RADIOISOTOPES; ENZYME ACTIVITY; ENZYMES; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Gluconacetobacter xylinus - enzymology; glucosides; Glucosyltransferases - isolation & purification; Glucosyltransferases - metabolism; GLYCOSYL TRANSFERASES; ISOTOPES; LABELLING; ligases; LIGHT NUCLEI; Macromolecular Substances; MEMBRANES; MICROORGANISMS; MOLECULAR WEIGHT; NUCLEI; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PEPTIDES; PHOSPHORUS 32; PHOSPHORUS ISOTOPES; Phosphorus Radioisotopes; POLYPEPTIDES; PROTEINS; RADIOISOTOPES; TRANSFERASES; uridine; uridine 5'-diphosphoglucose; Uridine Diphosphate Glucose - analogs & derivatives; Uridine Diphosphate Glucose - metabolism; Uridine Diphosphate Sugars - metabolism; Catalytic subunit; Ligand binding; Radiolabelling; Gel electrophoresis; Enzyme; Saturation; Binding site; Photoaffinity labelling; Subunit; Cellulose synthase (UDP-forming); Autoradiography; Bacteria; Ultraviolet visible spectrometry
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(19)34039-6

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.