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Domain folding and flexibility of Escherichia coli FtsZ determined by tryptophan site-directed mutagenesis.

ABSTRACT: FtsZ has two domains, the amino GTPase domain with a Rossmann fold, and the carboxyl domain that resembles the chorismate mutase fold. Bioinformatics analyses suggest that the interdomain interaction is stronger than the interaction of the protofilament longitudinal interfaces. Crystal B factor analysis of FtsZ and detected conformational changes suggest a connection between these domains. The unfolding/folding characteristics of each domain of FtsZ were tested by introducing tryptophans into the flexible region of the amino (F135W) and the carboxyl (F275W and I294W) domains. As a control, the mutation F40W was introduced in a more rigid part of the amino domain. These mutants showed a native-like structure with denaturation and renaturation curves similar to wild type. However, the I294W mutant showed a strong loss of functionality, both in vivo and in vitro when compared to the other mutants. The functionality was recovered with the double mutant I294W/F275A, which showed full in vivo complementation with a slight increment of in vitro GTPase activity with respect to the single mutant. The formation of a stabilizing aromatic interaction involving a stacking between the tryptophan introduced at position 294 and phenylalanine 275 could account for these results. Folding/unfolding of these mutants induced by guanidinium chloride was compatible with a mechanism in which both domains within the protein show the same stability during FtsZ denaturation and renaturation, probably because of strong interface interactions.

SUBMITTER: Diaz-Espinoza R

PROVIDER: S-EPMC2203363 | biostudies-literature | 2007 Aug

REPOSITORIES: biostudies-literature

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Domain folding and flexibility of Escherichia coli FtsZ determined by tryptophan site-directed mutagenesis.

Díaz-Espinoza Rodrigo R Garcés Andrea P AP Arbildua José J JJ Montecinos Felipe F Brunet Juan E JE Lagos Rosalba R Monasterio Octavio O

Protein science : a publication of the Protein Society 20070801 8

FtsZ has two domains, the amino GTPase domain with a Rossmann fold, and the carboxyl domain that resembles the chorismate mutase fold. Bioinformatics analyses suggest that the interdomain interaction is stronger than the interaction of the protofilament longitudinal interfaces. Crystal B factor analysis of FtsZ and detected conformational changes suggest a connection between these domains. The unfolding/folding characteristics of each domain of FtsZ were tested by introducing tryptophans into th ...[more]

PMID: 17656575

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Dataset Information

Domain folding and flexibility of Escherichia coli FtsZ determined by tryptophan site-directed mutagenesis.

Publications

Domain folding and flexibility of Escherichia coli FtsZ determined by tryptophan site-directed mutagenesis.

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