Site-specific and reversible anchoring of active proteins onto cellulose using a cellulosome-like complex

↓ Direkt till sidans innehåll
↓ Direkt till sidans sekundära innehåll (sidomenyn)

Search: onr:"swepub:oai:DiVA.org:kth-23499" > Site-specific and r...

1 of 1
Previous record
Next record
To hitlist

Site-specific and reversible anchoring of active proteins onto cellulose using a cellulosome-like complex

Eklund, Malin (author): KTH,Bioteknologi

Sandström, Kristofer (author): KTH,Bioteknologi

Teeri, Tuula (author): KTH,Bioteknologi

Nygren, Per-Åke (author): KTH,Bioteknologi

show less...

(creator_code:org_t)

Elsevier BV, 2004
2004
English.
In: Journal of Biotechnology. - : Elsevier BV. - 0168-1656 .- 1873-4863. ; 109:3, s. 277-286

Related links:: https://urn.kb.se/re...; show more...; https://doi.org/10.1...; show less...

Journal article (peer-reviewed)

Abstract Subject headings

Protein engineering strategies facilitating controlled and spontaneous assembly of macromolecular complexes are of great interest for the design of artificial multi-enzyme systems of pre-defined composition. Here we have combined affinity proteins from different sources to achieve specific and reversible anchoring of affinity domain-tagged reporter proteins to a cell ulose-anchored fusion protein. The design principle mimics the architecture of macromolecular cellulosome complexes produced by some cellulolytic microbes. A fusion protein between a cellulose-binding module (CBM1(Cel6A)) of the Trichoderma reesei cellobiohydrolase Cel6A and a five-domain staphylococcal protein A (SPA) was constructed to serve as platform for docking of easily detectable reporter proteins onto cellulose surfaces. In turn, the reporter proteins were produced as fusions to two copies of a SPA-binding affinity protein (an affibody denoted Z(SPA-1)), selected from a phage display library constructed by combinatorial protein engineering. In a series of experiments, involving repeated washing and low pH elution, affinity-tagged Enhanced Green Fluorescent Protein (EGFP) and Fusarium solani pisi lipase cutinase reporter proteins were both found to be specifically directed from solution to the same region of a cellulose filter paper where SPA-CBM1(Cel6A) fusion protein had been previously applied. This showed that the SPA-CBM1(Cel6A) fusion protein had been stably anchored to the cellulose surface without loss of binding capacity and that the interaction between SPA and the Z(SPA-1) affibody domains was selective. The generality of this biospecificity-driven system for assembly applications is discussed.

Subject headings

NATURVETENSKAP -- Biologi (hsv//swe)
NATURAL SCIENCES -- Biological Sciences (hsv//eng)

Keyword

cellulosome
assembly
affibody
affinity gene fusion
protein engineering
cellulose
bacterial receptor domain
binding domain
combinatorial library
degradation
expression
machines
affibody
chimeras
peptides
systems

Publication and Content Type

ref (subject category)
art (subject category)

Find in a library

Journal of Biotechnology (Search for host publication in LIBRIS)

To the university's database

1 of 1
Previous record
Next record
To hitlist

Find more in SwePub

By the author/editor: Eklund, Malin; Sandström, Krist ...; Teeri, Tuula; Nygren, Per-Åke

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...

Articles in the publication: Journal of Biote ...

By the university: Royal Institute of Technology

Search outside SwePub

Extend your search to:: Google; Google Book Search; Google Scholar

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.

LIBRIS.kb.se