dc.contributor.author |
Young, Paul |
en |
dc.contributor.author |
Kwon, H |
en |
dc.contributor.author |
Baker, Edward |
en |
dc.contributor.author |
Squire, Christopher |
en |
dc.coverage.spatial |
Canberra, Australia |
en |
dc.date.accessioned |
2016-08-09T22:55:13Z |
en |
dc.date.issued |
2016-04-04 |
en |
dc.identifier.citation |
Synthetic Biology Cutting Edge Symposium, Canberra, Australia, 04 Apr 2016 - 05 Apr 2016. Synthetic Biology Cutting Edge Symposium. 04 Apr 2016 |
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dc.identifier.uri |
http://hdl.handle.net/2292/29878 |
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dc.description.abstract |
Gram-positive bacterial pili and fimbriae are exposed to a hostile environment and must endure high tensile forces during the adhesion process. These ultra-stable proteins often contain internal crosslinks that impart crucial mechanical strength. Here, we report the first example of an isopeptide bond engineered de novo into an immunoglobulin-like protein domain. Through the site-directed mutagenesis of four residues we introduced isopeptide-forming residues into the hydrophobic core of FctB, an IgG-like minor pilin protein from Streptococcus pyogenes, to produce a covalent bond not present in the wild type protein. The isopeptide bond was engineered to covalently link the first and last β-strands, a position that is subjected to the highest mechanical stress. The presence of the engineered isopeptide-containing protein was confirmed by mass spectrometry and X-ray crystallography, and its thermal stability was measured as 81°C, an increase of 10°C over the wild type protein. This novel method for increasing the stability of IgG-like proteins has potential to be adopted by the field of antibody engineering, which share similar β-clasp Ig-type domains. |
en |
dc.relation.ispartof |
Synthetic Biology Cutting Edge Symposium |
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dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
en |
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
en |
dc.title |
Engineering protein stability with an intra-molecular isopeptide bond |
en |
dc.type |
Conference Item |
en |
pubs.author-url |
https://events.csiro.au/Events/2015/October/19/E-CUSYMP-160400 |
en |
pubs.finish-date |
2016-04-05 |
en |
pubs.start-date |
2016-04-04 |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Abstract |
en |
pubs.elements-id |
529093 |
en |
pubs.org-id |
Science |
en |
pubs.org-id |
Biological Sciences |
en |
pubs.org-id |
Science Research |
en |
pubs.org-id |
Maurice Wilkins Centre (2010-2014) |
en |
pubs.record-created-at-source-date |
2016-05-31 |
en |