Semi-synthetic Rab proteins as tools for studying intermolecular interactions.
Rab GTPases play a key role in the regulation of membrane traffic. Posttranslational geranylgeranylation is critical for their biological activity and is conferred by a Rab geranylgeranyl transferase (RabGGTase). To study the interactions between Rab proteins and RabGGTase, we used in vitro ligation methodology to generate a fluorescent semi-synthetic Rab7 protein. The obtained protein was functionally active and was used to demonstrate a micromolar affinity interaction of Rab7 with the RabGGTase in the absence of Rab escort protein (REP). This finding is consistent with an earlier proposed model according to which RabGGTase possesses two independent weak binding sites for REP and Rab proteins.
Iakovenko
A
A
Department of Physical Biochemistry, Max-Planck Institute for Molecular Physiology, Otto-Hahn-Strasse 11, 44227, Dortmund, Germany.
Rostkova
E
E
Merzlyak
E
E
Hillebrand
A M
AM
Thomä
N H
NH
Goody
R S
RS
Alexandrov
K
K
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D004735:Energy Transfer; D004926:Escherichia coli; D007700:Kinetics; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D020691:rab GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D000090802:rab7 GTP-Binding Proteins
D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; D000090802:rab7 GTP-Binding Proteins; D019883:Alkyl and Aryl Transferases; C075607:Rab geranylgeranyltransferase; D020691:rab GTP-Binding Proteins