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

Lookup NU author(s): Dr Benjamin Chaffey, Elizabeth Mitchell, Dr Mark Birch, Professor Jeremy LakeyORCiD

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Abstract

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.


Publication metadata

Author(s): Chaffey BT, Mitchell E, Birch MA, Lakey JH

Publication type: Article

Publication status: Published

Journal: International Journal of Nanomedicine

Year: 2008

Volume: 3

Issue: 3

Pages: 287-293

Print publication date: 01/01/2008

ISSN (print): 1176-9114

ISSN (electronic): 1178-2013

Publisher: Dove Medical Press Ltd


Funding

Funder referenceFunder name
13/E19051BBSRC

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