Properties of the subunits of wheat germ initiation factor 3.
Wheat germ initiation factor 3 (eukaryotic initiation factor 3, eIF-3) contains ten non-identical subunits (p116, p107, p87, p83, p56, p45, p41, p36, p34 and p28). Monoclonal antibodies to all except two of the subunits (p41 and p28) were obtained. None of the monoclonal antibodies react with more than one subunit, and only monoclonal antibodies to p36 inhibit the ability of eIF-3 to support initiation of polypeptide synthesis. Two of the subunits (p116 and p107) are highly basic polypeptides (pI greater than or equal to 8); five (p87, p56, p45, p34 and p28) are acidic polypeptides (pI = 5.4-6.1); and three (p83, p41 and p36) appear to exist in more than one isoelectric form. Eight of the subunits of eIF-3 are iodinated rapidly in vitro; the highest incorporation is into p56 and the lowest incorporation is into p28. No incorporation into p41 or p28 is observed. When eIF-3 is treated with N-[3H]ethylmaleimide, approx. 30 alkyl groups per eIF-3 are incorporated, and the eIF-3 is inactivated. No incorporation into p83 or p28 is observed; incorporation of the alkyl groups into the other eight subunits occurs at different rates. The rate of inactivation of eIF-3 by N-ethylmaleimide is slower than the overall rate of incorporation of alkyl groups. eIF-3 is stable between pH 5.5 and 10. Below pH 5.5, eIF-3 is inactivated and precipitation of protein occurs. Partial dissociation of the subunits and inactivation of eIF-3 is obtained by treatment with 2 M urea. Attempts to reassociate the subunits into an active particle were unsuccessful.
Heufler
C
C
Department of Chemistry, University of Texas, Austin 78712.
Browning
K S
KS
Ravel
J M
JM
D000478:Alkylation; D000911:Antibodies, Monoclonal; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide; D039621:Eukaryotic Initiation Factor-3; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007457:Iodine Radioisotopes; D007526:Isoelectric Point; D046911:Macromolecular Substances; D010452:Peptide Biosynthesis; D010448:Peptide Initiation Factors / Q000032:analysis / Q000378:metabolism*; D010944:Plants / Q000032:analysis*; D011489:Protein Denaturation; D014908:Triticum; D014508:Urea / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D039621:Eukaryotic Initiation Factor-3; D007457:Iodine Radioisotopes; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D014508:Urea; D005033:Ethylmaleimide