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Characterization of cyanogen bromide peptides of hypoxanthine phosphoribosyltransferase by a micromethod. [/sup 125/I, /sup 3/H, /sup 35/S tracer techniques]

Journal Article · · Anal. Biochem.; (United States)
Human hypoxanthine phosphoribosyltransferase (HPRT-EC 2.4.2.8), either from erythrocytes labeled in vitro with /sup 125/I or from HeLa cells labeled in vivo with (/sup 3/H)leucine, (/sup 3/H)lysine, or (/sup 3/H)arginine, was cleaved with cyanogen bromide. All four labeled enzymes produced cyanogen bromide peptides with molecular weights of 7800, 5600, 4100, 2500, and 1300 which were separated by sodium dodecyl sulfate urea polyacrylamide gel electrophoresis. Experiments with (/sup 35/S)methionine-labeled HPRT indicated that 88 to 94% of the /sup 35/S label was released by the cyanogen bromide reaction. The relative distribution of radioactivity in each of the tritiated peptides indicates that there are 20 leucines, 15 lysines, and 12 arginines per enzyme subunit.
Research Organization:
Univ. of California, Berkeley
OSTI ID:
7294629
Journal Information:
Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 82:1; ISSN ANBCA
Country of Publication:
United States
Language:
English

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