GTP Hydrolysis of Cell Division Protein FtsZ:  Evidence that the Active Site Is Formed by the Association of Monomers

The essential prokaryotic cell division protein FtsZ is a tubulin homologue that forms a ring at the division site. FtsZ forms polymers in a GTP-dependent manner. Recent biochemical evidence has shown that FtsZ forms multimeric structures in vitro and in vivo and functions as a self-activating GTPas...

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Bibliographic Details
Published in:Biochemistry (Easton) 2002-01, Vol.41 (2), p.521-529
Main Authors: Scheffers, Dirk-Jan, de Wit, Janny G, den Blaauwen, Tanneke, Driessen, Arnold J. M
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacterial Proteins - chemistry
Binding Sites
Calcium - pharmacology
Cysteine - chemistry
Cytoskeletal Proteins
Dose-Response Relationship, Drug
Escherichia coli - enzymology
Glutamic Acid - chemistry
GTP Phosphohydrolases - metabolism
Guanosine Triphosphate - metabolism
Hydrolysis
Light
Magnesium - pharmacology
Microscopy, Electron
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Mutation
Protein Binding
Protein Conformation
Protein Structure, Tertiary
Scattering, Radiation
Sequence Homology, Amino Acid
Time Factors
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Summary:The essential prokaryotic cell division protein FtsZ is a tubulin homologue that forms a ring at the division site. FtsZ forms polymers in a GTP-dependent manner. Recent biochemical evidence has shown that FtsZ forms multimeric structures in vitro and in vivo and functions as a self-activating GTPase. Structural analysis of FtsZ points to an important role for the highly conserved tubulin-like loop 7 (T7-loop) in the self-activation of GTP hydrolysis. The T7-loop was postulated to form the active site together with the nucleotide-binding site on an adjacent FtsZ monomer. To characterize the role of the T7-loop of Escherichia coli FtsZ, we have mutagenized residues M206, N207, D209, D212, and R214. All the mutant proteins, except the R214 mutant, are severely affected in polymerization and GTP hydrolysis. Charged residues D209 and D212 cannot be substituted with a glutamate residue. All mutants interact with wild-type FtsZ in vitro, indicating that the T7-loop mutations do not abolish FtsZ self-association. Strikingly, in mixtures of wild-type and mutant proteins, most mutants are capable of inhibiting wild-type GTP hydrolysis. We conclude that the T7-loop is part of the active site for GTP hydrolysis, formed by the association of two FtsZ monomers.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi011370i