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Conformational changes of FtsZ reported by tryptophan mutants.

dc.contributor.author Chen, Yaodong
dc.contributor.author Erickson, Harold Paul
dc.date.accessioned 2018-04-01T14:58:57Z
dc.date.available 2018-04-01T14:58:57Z
dc.date.issued 2011-05-03
dc.identifier.issn 0006-2960
dc.identifier.issn 1520-4995
dc.identifier.uri https://hdl.handle.net/10161/16460
dc.description.abstract E. coli FtsZ has no native tryptophan. We showed previously that the mutant FtsZ L68W gave a 2.5-fold increase in trp fluorescence when assembly was induced by GTP. L68 is probably buried in the protofilament interface upon assembly, causing the fluorescence increase. In the present study we introduced trp residues at several other locations and examined them for assembly-induced fluorescence changes. L189W, located on helix H7 and buried between the N- and C-terminal subdomains, showed a large fluorescence increase, comparable to L68W. This may reflect a shift or rotation of the two subdomains relative to each other. L160W showed a smaller increase in fluorescence, and Y222W a decrease in fluorescence, upon assembly. These two are located on the surface of the N and C subdomains, near the domain boundary. The changes in fluorescence may reflect movements of the domains or of nearby side chains. We prepared a double mutant Y222W/S151C and coupled ATTO-655 to the cys. The Cα of trp in the C-terminal subdomain was 10 Å away from that of the cys in the N-terminal subdomain, permitting the ATTO to make van der Waals contact with the trp. The ATTO fluorescence showed strong tryptophan-induced quenching. The quenching was reduced following assembly, consistent with a movement apart of the two subdomains. Movements of one to several angstroms are probably sufficient to account for the changes in trp fluorescence and trp-induced quenching of ATTO. Assembly in GDP plus DEAE dextran produces tubular polymers that are related to the highly curved, mini-ring conformation. No change in trp fluorescence was observed upon assembly of these tubes, suggesting that the mini-ring conformation is the same as that of a relaxed, monomeric FtsZ.
dc.language eng
dc.relation.ispartof Biochemistry
dc.relation.isversionof 10.1021/bi200106d
dc.subject Acrylamide
dc.subject Tryptophan
dc.subject Bacterial Proteins
dc.subject Cytoskeletal Proteins
dc.subject Microscopy, Electron
dc.subject Spectrometry, Fluorescence
dc.subject Protein Conformation
dc.subject Mutation
dc.subject Models, Molecular
dc.title Conformational changes of FtsZ reported by tryptophan mutants.
dc.type Journal article
dc.date.updated 2018-04-01T14:58:57Z
pubs.issue 21
pubs.organisational-group School of Medicine
pubs.organisational-group Duke
pubs.organisational-group Duke Cancer Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Biochemistry
pubs.organisational-group Basic Science Departments
pubs.organisational-group Cell Biology
pubs.publication-status Published
pubs.volume 50


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