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Photoaffinity labeling of the messenger RNA cap-binding protein

Journal Article · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5096717
A photoaffinity derivative of a cap analogue, ..gamma..(/sup 32/P)-(4-(benzoylphenyl)methylamido)-7-methylguanosine 5'-triphosphate, was reacted with the 28-kDa messenger RNA cap-binding protein (CBP), purified by affinity chromatography from rabbit erythrocyte lysate. The predominant labeling was of the 28-kDa species. No labeling occurred without photoillumination or in the presence of 250 ..mu..M m/sup 7/GTP. In addition CBP, two polypeptides of 19 and 17kDa, present in the affinity-purified preparation, were specifically labeled. Another sample tested was the eIF-3 region of a sucrose gradient fractionation, in 100 mM KCl, of rabbit reticulocyte high salt ribosomal wash. The only polypeptide labeled was the 28-kDa CBP. Labeling was more intense than with an equivalent amount of affinity-purified CBP, suggesting the presence of stabilizing or stimulatory factors in the eIF-3 preparation.
Research Organization:
Univ. of Kentucky, Lexington
OSTI ID:
5096717
Report Number(s):
CONF-8606151-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
Country of Publication:
United States
Language:
English