title: Polyisoprenylation of the CAAX motif--an in vitro protein synthesis study.
abstract: A number of proteins, including most nuclear lamins, certain fungal mating pheromones, G-protein gamma-subunits and ras proteins, contain a C-terminal cysteine-aliphatic-aliphatic-undefined amino acid (CAAX) motif which is thought to be a roughly defined consensus sequence capable of directing a series of posttranslational events, beginning with the addition of a polyisoprene moiety to the cysteine. So far such a motif has been found in every protein known to have this type of modification. We have utilized the rabbit reticulocyte lysate translation system, which is capable of carrying out the polyisoprene modification in vitro, to investigate features of the C-terminal motif which affect its suitability as a substrate. We demonstrate that a cysteine is only isoprenylated when situated at position -4 from the C-terminus. We further show that the presence of a glycine at position -3 or a terminal aromatic residue, features typical of some G-protein alpha subunits, cause a reduction and abolition respectively of isoprenylation.
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Newman
authors.forename: E
authors.initials: E
authors.lastname: Kube
authors.forename: V
authors.initials: V
authors.lastname: Gerke
authors.forename: K
authors.initials: K
authors.lastname: Weber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010779:Photofluorography; D010957:Plasmids; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D012717:Sesquiterpenes; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
chemical_list: D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate