dc.contributor.author |
Rechiche, Mohamed Othman |
en |
dc.contributor.author |
Plowman, JE |
en |
dc.contributor.author |
Harland, DP |
en |
dc.contributor.author |
Lee, Tet |
en |
dc.contributor.author |
Lott, Jeremy |
en |
dc.date.accessioned |
2018-10-28T19:39:30Z |
en |
dc.date.issued |
2018-06 |
en |
dc.identifier.citation |
Protein Expression and Purification 146:34-44 Jun 2018 |
en |
dc.identifier.issn |
1046-5928 |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/43637 |
en |
dc.description.abstract |
Keratin-associated proteins (KAPs) were identified 70 years ago in wool follicles. KAPs are encoded by several multi-gene families and are classified into three different groups: ultra-high sulfur (UHS), high sulfur (HS) and high glycine-tyrosine (HGT). KAPs are the major constituent of the matrix between the hair keratin intermediate filaments (IFs), and stabilise hair structure by extensive disulfide bonding. However, detailed molecular structural information is lacking for KAPs and for KAP interactions with IFs. As a preliminary step towards their biophysical and structural characterization, we have expressed and purified a HS KAP (KAP11.1) and a HGT KAP (KAP6.1). The expression and purification of KAPs is challenging because they are cysteine-rich proteins with unusual amino acid compositions, they tend to be insoluble in isolation and are prone to forming aggregates in solution. Here we describe the high yield production of pure, soluble KAPs in a chaotrope- and detergent-free buffer. This method has the potential to be used for the overproduction of other KAPs and similar cysteine-rich proteins with high isoelectric points. |
en |
dc.format.medium |
Print-Electronic |
en |
dc.language |
eng |
en |
dc.publisher |
Elsevier |
en |
dc.relation.ispartofseries |
Protein Expression and Purification |
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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.rights.uri |
http://creativecommons.org/licenses/by-nc-nd/4.0 |
en |
dc.subject |
Hair |
en |
dc.subject |
Humans |
en |
dc.subject |
Escherichia coli |
en |
dc.subject |
Buffers |
en |
dc.subject |
Chromatography, Affinity |
en |
dc.subject |
Chromatography, Gel |
en |
dc.subject |
Cloning, Molecular |
en |
dc.subject |
Amino Acid Sequence |
en |
dc.subject |
Protein Denaturation |
en |
dc.subject |
Keratins |
en |
dc.subject |
Protein Stability |
en |
dc.title |
Expression and purification of high sulfur and high glycine-tyrosine keratin-associated proteins (KAPs) for biochemical and biophysical characterization |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1016/j.pep.2017.12.006 |
en |
pubs.begin-page |
34 |
en |
pubs.volume |
146 |
en |
dc.rights.holder |
Copyright: Elsevier |
en |
dc.identifier.pmid |
29391282 |
en |
pubs.end-page |
44 |
en |
pubs.publication-status |
Published |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
Article |
en |
pubs.elements-id |
723019 |
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 |
dc.identifier.eissn |
1096-0279 |
en |
pubs.record-created-at-source-date |
2019-07-08 |
en |
pubs.dimensions-id |
29391282 |
en |