English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Enhancing recombinant protein solubility with ubiquitin-like small archeal modifying protein fusion partners

Varga, S., Pathare, G. R., Baka, E., Boicu, M., Kriszt, B., Szekacs, A., et al. (2015). Enhancing recombinant protein solubility with ubiquitin-like small archeal modifying protein fusion partners. JOURNAL OF MICROBIOLOGICAL METHODS, 118, 113-122. doi:10.1016/j.mimet.2015.08.017.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-5C4F-0 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-5C50-A
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Varga, Sandor1, Author
Pathare, Ganesh Ramnath2, Author              
Baka, Erzsebet1, Author
Boicu, Marius2, Author              
Kriszt, Balazs1, Author
Szekacs, Andras1, Author
Zinzula, Luca2, Author              
Kukolya, Jozsef1, Author
Nagy, Istvan2, Author              
Affiliations:
1external, ou_persistent22              
2Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

Content

show
hide
Free keywords: EXPRESSION; TAGSProtein expression; Protein solubility; Protein purification; SAMP; Tag removal; E.coli;
 Abstract: A variety of protein expression tags with different biochemical properties has been used to enhance the yield and solubility of recombinant proteins. Ubiquitin, SUMO (small ubiquitin-like modifier) and prokaryotic ubiquitin like MoaD (molybdopterin synthase, small subunit) fusion tags are getting more popular because of their small size. In this paper we report on the use of ubiquitin-like small archaeal modifier proteins (SAMPs) as fusion tags since they proved to increase expression yield, stability and solubility in our experiments. Equally important, they did not co-purify with proteins of the expression host and there was information that their specific JAB1/MPN/Mov34 metalloenzyme (JAMM) protease can recognize the C-terminal VSGG sequence when SAMPs fused, either branched or linearly to target proteins, and cleave it specifically. SAMPs and JAMM proteases from Haloferax volcanii, Thermoplasma acidophilum, Methanococcoides burtonii and Nitrosopumilus maritimus were selected, cloned, expressed heterologously in Escherichia coli and tested as fusion tags and cleaving proteases, respectively. Investigated SAMPs enhanced protein expression and solubility on a wide scale. T. acidophilum SAMPs Ta0895 and Ta01019 were the best performing tags and their effect was comparable to the widely used maltose binding protein (MBP) and N utilization substance protein A (NusA) tags. Moreover, H. volcanii SAMP Hvo_2619 contribution was mediocre, whereas M. burtonii Mbur_1415 could not be expressed. Out of four investigated JAMM proteases, only Hvo_2505 could cleave fusion tags. Interestingly, it was found active not only on its own partner substrate Hvo_2619, but it also cleaved off Ta0895. (C) 2015 Elsevier B.V. All rights reserved.

Details

show
hide
Language(s): eng - English
 Dates: 2015
 Publication Status: Published in print
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: JOURNAL OF MICROBIOLOGICAL METHODS
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS : ELSEVIER SCIENCE BV
Pages: - Volume / Issue: 118 Sequence Number: - Start / End Page: 113 - 122 Identifier: ISSN: 0167-7012