Amino acid sequence of rubber elongation factor protein associated with rubber particles in Hevea latex.
The amino acid sequence of rubber elongation factor, a recently discovered protein tightly bound to rubber particles isolated from the commercial rubber tree Hevea brasiliensis, is presented. The role of this protein in rubber elongation and its interaction with prenyltransferase and rubber particles have been discussed in the preceding paper in this series (Dennis, M. S., and Light, D. R. (1989) J. Biol. Chem. 264, 18608-18617). Trypsin, Staphylococcus protease, chymotrypsin, acetic acid, and hydroxylamine cleavage were used to generate peptide fragments that were isolated by reverse phase high pressure liquid chromatography and analyzed by amino acid composition and automated Edman degradation. Each digest contained one blocked peptide identified as the amino terminus. The blocked amino-terminal peptide from the tryptic digest was analyzed by amino acid composition, fast atom bombardment mass spectrometry (molecular ion 1659.9), subdigested with Staphylococcus protease for partial sequence analysis, and finally deblocked with bovine liver acyl-peptide hydrolase removing an acetylalanine to allow analysis by Edman degradation. Rubber elongation factor is 137 amino acids long, has a molecular mass of 14,600 daltons, and lacks four amino acids: cysteine, methionine, histidine, and tryptophan. The NH2 terminus is highly charged and contains only acidic residues (5 of the first 12 amino acids). The first four amino acids are highly represented in other known NH2-terminally acetylated proteins. Comparison of the sequence of rubber elongation factor with other known sequences does not reveal significant sequence similarities that would suggest an evolutionary relationship.
Dennis
M S
MS
Department of Medicinal Biology, Genentech, Inc., South San Francisco, California 94080.
Henzel
W J
WJ
Bell
J
J
Kohr
W
W
Light
D R
DR
D000085:Acetates; D019342:Acetic Acid; D000485:Allergens; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D052179:Antigens, Plant; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D019811:Hydroxylamine; D006898:Hydroxylamines; D007840:Latex / Q000032:analysis*; D013058:Mass Spectrometry; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010940:Plant Proteins; D010944:Plants / Q000032:analysis*; D011506:Proteins / Q000302:isolation & purification; D014197:Trees; D014357:Trypsin
D000085:Acetates; D000485:Allergens; D000596:Amino Acids; D052179:Antigens, Plant; D006898:Hydroxylamines; D007840:Latex; D010446:Peptide Fragments; D010940:Plant Proteins; D011506:Proteins; C061477:REF protein, Hevea brasiliensis; D019811:Hydroxylamine; D002918:Chymotrypsin; D014357:Trypsin; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D019342:Acetic Acid