Photoaffinity labeling of the messenger RNA cap-binding protein

Journal Article · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5096717

Rhoads, R; Patzelt, E; Blaas, D; Kuechler, E

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), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
LABELLING
AFFINITY
MESSENGER-RNA
PHOSPHATES
PHOSPHORUS 32
TRACER TECHNIQUES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RNA
550201* - Biochemistry- Tracer Techniques