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Monoclonal antibodies against eIF-2B inhibit translation in rabbit reticulocyte lysate

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6079488
; ;

eIF-2B is a five-subunit 280 kDa protein whose function is to recycle inactive translation factor eIF-2:GDP to its active eIF-2:GTP form through GDP-GTP exchange. To understand the function of individual eIF-2B subunits, they have recently prepared F1, a mouse MAb against highly-purified rabbit eIF-2B. Western analysis shows that F1 recognizes the 82 kDa subunit (epsilon) of eIF-2B either free or in the eIF-2 x 2B complex. F1 inhibits incorporation of /sup 3/H-leucine in hemin-supplemented rabbit reticulocyte lysate with biphasic kinetics with the concomitant loss of polysomes. This inhibition is accompanied by a six to eight-fold decrease in the amount of met-tRNAi in 43S pre-initiation complexes and the appearance of a novel 15S complex. The phosphorylation state of eIF-2 is not altered during F1-dependent translational inhibition. Addition of purified eIF-2B efficiently rescues translation, with reformation of polysomes. eIF-2B catalyzes displacement /sup 3/H-GDP from the /sup 3/H-GDP x eIF-2 binary complex with either GDP or GTP, and this exchange is not inhibited by F1. This suggests an alternative mechanism of inhibition and/or role for eIF-2B in the formation of the eIF-2 x met-tRNAi x GTP ternary complex.

Research Organization:
National Institute of Health, Bethesda, MD
OSTI ID:
6079488
Report Number(s):
CONF-870644-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
ANTIBODIES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
ERYTHROCYTES
FUNCTIONS
GUANYLIC ACID
INHIBITION
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LEUCINE
MAMMALS
MATERIALS
MICE
MONOCLONAL ANTIBODIES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHOSPHORYLATION
PROTEINS
RABBITS
REACTION KINETICS
RETICULOCYTES
RODENTS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES