title: Chemical biology of protein lipidation: semi-synthesis and structure elucidation of prenylated RabGTPases.
abstract: Rab/Ypt guanosine triphosphatases (GTPases) represent a family of key membrane traffic regulators in eukaryotic cells. For their function Rab/Ypt proteins require double modification with two covalently bound geranylgeranyl lipid moieties at the C-terminus. Generally, prenylated proteins are very difficult to obtain by recombinant or enzymatic methods. We generated prenylated RabGTPases using a combination of chemical synthesis and protein engineering. This semi-synthesis depends largely on the availability of functionalized prenylated peptides corresponding to the proteins' native structure or modifications. We developed solution phase and solid phase strategies for the generation of peptides corresponding to the prenylated C-terminus of Rab7 GTPase in preparative amounts enabling us to crystallize the mono-prenylated Ypt1:RabGDI complex. The structure of the complex provides a structural basis for the ability of RabGDI to inhibit the release of nucleotide by Rab proteins and a molecular basis for understanding a RabGDI mutant that causes mental retardation in humans.
authors.affiliation: Max-Planck-Institut für Molekulare Physiologie, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Watzke
authors.forename: Luc
authors.initials: L
authors.lastname: Brunsveld
authors.forename: Thomas
authors.initials: T
authors.lastname: Durek
authors.forename: Kirill
authors.initials: K
authors.lastname: Alexandrov
authors.forename: Alexey
authors.initials: A
authors.lastname: Rak
authors.forename: Roger S
authors.initials: RS
authors.lastname: Goody
authors.forename: Herbert
authors.initials: H
authors.lastname: Waldmann
mesh_terms: D008958:Models, Molecular; D011487:Protein Conformation; D015202:Protein Engineering; D017368:Protein Prenylation; D020691:rab GTP-Binding Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D020691:rab GTP-Binding Proteins