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PMID: 2123345 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Posttranslationally processed structure of the human platelet protein smg p21B: evidence for geranylgeranylation and carboxyl methylation of the C-terminal cysteine.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 22 ·1990-11-00 ·Pages 8960-4

Kawata M, Farnsworth CC, Yoshida Y, Gelb MH, Glomset JA, Takai Y

Abstract

smg p21A and -B are small GTP-binding proteins that share putative effector and consensus C-terminal sequences with ras p21 proteins. In the present report, we showed that human platelet smg p21B became labeled when intact platelets were incubated with exogenous [3H]mevalonolactone and when a purified preparation of smg p21B was incubated with bovine brain membranes and S-adenosyl-L-[methyl-3H]methionine. In addition, we demonstrated by gas chromatography/mass spectrometry that treatment of smg p21B with Raney nickel released a geranylgeranyl moiety in a molar ratio of about 1:1. In contrast, treatment of smg p21B with NH2OH or KOH yielded no evidence for the presence of a palmitoyl thioester. Extensive digestion of smg p21B with Achromobacter protease I yielded two C-terminal tripeptides that contained serine and cysteine in a molar ratio of 2:1. Both peptides were modified by a thioether-linked geranylgeranyl group. One of the peptides comigrated with a 3H-labeled proteolytic product of methylated smg p21B on reverse-phase HPLC and this peptide appeared at the same retention time as that of the other peptide after being treated with KOH. Since the cDNA-predicted C-terminal sequence of smg p21B contains a unique Ser-Ser-Cys peptide within its C-terminal domain, -Lys-Lys-Ser-Ser-Cys-Gln-Leu-Leu184, these results indicate that smg p21B is posttranslationally modified by geranylgeranylation of Cys-181 and suggest that further modifications cause proteolytic removal of the three predicted C-terminal amino acids followed by partial methylation of the cysteinyl carboxyl group.

MeSH Terms
Blood Platelets/metabolism Cysteine/metabolism GTP-Binding Proteins/chemistry,metabolism Gas Chromatography-Mass Spectrometry Humans In Vitro Techniques Methylation Mevalonic Acid/analogs & derivatives,chemistry Polyisoprenyl Phosphates/metabolism Protein Processing, Post-Translational rap GTP-Binding Proteins
Chemicals
Polyisoprenyl Phosphates mevalonolactone GTP-Binding Proteins rap GTP-Binding Proteins Cysteine Mevalonic Acid
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kawata M
Department of Biochemistry, Kobe University School of Medicine, Japan.
Farnsworth C C
Yoshida Y
Gelb M H
Glomset J A
Takai Y
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.22.8960
Published
1990-11-00
Pages
8960-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55080
Subset
IM
Grants
NIGMS NIH HHS · GM00562 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.22.8960 PMC Full Text View journal details
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