A generic expression system to produce proteins that co-assemble with alkane thiol SAM

Benjamin T Chaffey1, Elizabeth Mitchell1, Mark A Birch2, Jeremy H Lakey11The Institute for Cell and Molecular Biosciences; 2The School of Surgical and Reproductive Sciences, The Medical School, Framlington Place, The University of Newcastle-upon-Tyne, Newcastle-upon-Tyne, Great BritainAbstract: Surf...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Benjamin T Chaffey, Elizabeth Mitchell, Mark A Birch, Jeremy H Lakey
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2008
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/c5bf1697c942432fa37eb413acaae43b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:c5bf1697c942432fa37eb413acaae43b
record_format	dspace
spelling	oai:doaj.org-article:c5bf1697c942432fa37eb413acaae43b2021-12-02T05:09:47ZA generic expression system to produce proteins that co-assemble with alkane thiol SAM1176-91141178-2013https://doaj.org/article/c5bf1697c942432fa37eb413acaae43b2008-10-01T00:00:00Zhttp://www.dovepress.com/a-generic-expression-system-to-produce-proteins-that-co-assemble-with--a2425https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Benjamin T Chaffey1, Elizabeth Mitchell1, Mark A Birch2, Jeremy H Lakey11The Institute for Cell and Molecular Biosciences; 2The School of Surgical and Reproductive Sciences, The Medical School, Framlington Place, The University of Newcastle-upon-Tyne, Newcastle-upon-Tyne, Great BritainAbstract: Surface biology aims to observe and control biological processes by combining bio-, surface, and physical chemistry. Self-assembled monolayers (SAM) on gold surfaces have provided excellent methods for nanoscale surface preparation for such studies. However, extension of this work requires the specific immobilization of whole protein domains and the direct incorporation of recombinant proteins into SAM is still problematic. In this study a short random coil peptide has been designed to insert into thioalkane layers by formation of a hydrophobic helix. Surface plasmon resonance (SPR) studies show that specific immobilization via the internal cysteine is achieved. Addition of the peptide sequence to the terminus of a protein at the genetic level enables the production of a range of recombinant fusion-proteins with good yield. SPR shows that the proteins display the same gold-binding behavior as the peptide. It is shown that cell growth control can be achieved by printing the proteins using soft lithography with subsequent infilling with thio-alkanes The expression plasmid is constructed so that any stable protein domain can be easily cloned, expressed, purified and immobilized.Keywords: self-assembling monolayer, protein immobilization, nanotechnology, circular dichroism spectroscopy, surface plasmon resonance, osteoblast Benjamin T ChaffeyElizabeth MitchellMark A BirchJeremy H LakeyDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2008, Iss Issue 3, Pp 287-293 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Benjamin T Chaffey Elizabeth Mitchell Mark A Birch Jeremy H Lakey A generic expression system to produce proteins that co-assemble with alkane thiol SAM
description	Benjamin T Chaffey1, Elizabeth Mitchell1, Mark A Birch2, Jeremy H Lakey11The Institute for Cell and Molecular Biosciences; 2The School of Surgical and Reproductive Sciences, The Medical School, Framlington Place, The University of Newcastle-upon-Tyne, Newcastle-upon-Tyne, Great BritainAbstract: Surface biology aims to observe and control biological processes by combining bio-, surface, and physical chemistry. Self-assembled monolayers (SAM) on gold surfaces have provided excellent methods for nanoscale surface preparation for such studies. However, extension of this work requires the specific immobilization of whole protein domains and the direct incorporation of recombinant proteins into SAM is still problematic. In this study a short random coil peptide has been designed to insert into thioalkane layers by formation of a hydrophobic helix. Surface plasmon resonance (SPR) studies show that specific immobilization via the internal cysteine is achieved. Addition of the peptide sequence to the terminus of a protein at the genetic level enables the production of a range of recombinant fusion-proteins with good yield. SPR shows that the proteins display the same gold-binding behavior as the peptide. It is shown that cell growth control can be achieved by printing the proteins using soft lithography with subsequent infilling with thio-alkanes The expression plasmid is constructed so that any stable protein domain can be easily cloned, expressed, purified and immobilized.Keywords: self-assembling monolayer, protein immobilization, nanotechnology, circular dichroism spectroscopy, surface plasmon resonance, osteoblast
format	article
author	Benjamin T Chaffey Elizabeth Mitchell Mark A Birch Jeremy H Lakey
author_facet	Benjamin T Chaffey Elizabeth Mitchell Mark A Birch Jeremy H Lakey
author_sort	Benjamin T Chaffey
title	A generic expression system to produce proteins that co-assemble with alkane thiol SAM
title_short	A generic expression system to produce proteins that co-assemble with alkane thiol SAM
title_full	A generic expression system to produce proteins that co-assemble with alkane thiol SAM
title_fullStr	A generic expression system to produce proteins that co-assemble with alkane thiol SAM
title_full_unstemmed	A generic expression system to produce proteins that co-assemble with alkane thiol SAM
title_sort	generic expression system to produce proteins that co-assemble with alkane thiol sam
publisher	Dove Medical Press
publishDate	2008
url	https://doaj.org/article/c5bf1697c942432fa37eb413acaae43b
work_keys_str_mv	AT benjamintchaffey agenericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT elizabethmitchell agenericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT markabirch agenericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT jeremyhlakey agenericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT benjamintchaffey genericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT elizabethmitchell genericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT markabirch genericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam AT jeremyhlakey genericexpressionsystemtoproduceproteinsthatcoassemblewithalkanethiolsam
_version_	1718400558230929408

A generic expression system to produce proteins that co-assemble with alkane thiol SAM

Ejemplares similares