In vitro association of fragments of a beta-sheet membrane protein

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Standard

In vitro association of fragments of a beta-sheet membrane protein. / Debnath, D; Nielsen, K L; Otzen, D E.

I: Advances in Biophysical Chemistry, Bind 148, Nr. 1-3, 01.05.2010, s. 112-20.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Debnath, D, Nielsen, KL & Otzen, DE 2010, 'In vitro association of fragments of a beta-sheet membrane protein', Advances in Biophysical Chemistry, bind 148, nr. 1-3, s. 112-20. https://doi.org/10.1016/j.bpc.2010.03.004

APA

Debnath, D., Nielsen, K. L., & Otzen, D. E. (2010). In vitro association of fragments of a beta-sheet membrane protein. Advances in Biophysical Chemistry, 148(1-3), 112-20. https://doi.org/10.1016/j.bpc.2010.03.004

CBE

Debnath D, Nielsen KL, Otzen DE. 2010. In vitro association of fragments of a beta-sheet membrane protein. Advances in Biophysical Chemistry. 148(1-3):112-20. https://doi.org/10.1016/j.bpc.2010.03.004

MLA

Debnath, D, K L Nielsen, og D E Otzen. "In vitro association of fragments of a beta-sheet membrane protein". Advances in Biophysical Chemistry. 2010, 148(1-3). 112-20. https://doi.org/10.1016/j.bpc.2010.03.004

Vancouver

Debnath D, Nielsen KL, Otzen DE. In vitro association of fragments of a beta-sheet membrane protein. Advances in Biophysical Chemistry. 2010 maj 1;148(1-3):112-20. https://doi.org/10.1016/j.bpc.2010.03.004

Author

Debnath, D ; Nielsen, K L ; Otzen, D E. / In vitro association of fragments of a beta-sheet membrane protein. I: Advances in Biophysical Chemistry. 2010 ; Bind 148, Nr. 1-3. s. 112-20.

Bibtex

@article{4148bad3e8e647bf973e56075e7ecc34,
title = "In vitro association of fragments of a beta-sheet membrane protein",
abstract = "Although the beta-barrel membrane protein OmpA can be produced in a biologically active form in E. coli from co-expressed fragments, the fragments have not been demonstrated to associate in vitro. We have produced 3 complementary fragment pairs of OmpA which can associate to form a folded complex according to the SDS band-shift assay. We are able to convert 25-35% of the fragment populations to non-covalent but SDS-stable complexes. The periplasmic chaperone Skp effectively prevented this association. Two separately expressed and purified overlapping fragments of OmpA can form a protease-resistant complex that undergoes the characteristic band-shift upon heating. Our work demonstrates that although membrane insertion and folding of beta-barrel membrane proteins may be a cooperative process, the fragments can associate in vitro without any additional components. However, the low yield and slow folding rates indicate that partially unfolded or destabilized beta-sheet membrane proteins can potentially engage in many non-native interactions.",
keywords = "Amino Acid Sequence, Bacterial Outer Membrane Proteins, DNA-Binding Proteins, Escherichia coli Proteins, Hot Temperature, Kinetics, Models, Molecular, Molecular Chaperones, Peptide Hydrolases, Protein Folding, Protein Structure, Secondary, Protein Structure, Tertiary, Substrate Specificity",
author = "D Debnath and Nielsen, {K L} and Otzen, {D E}",
year = "2010",
month = may,
day = "1",
doi = "10.1016/j.bpc.2010.03.004",
language = "English",
volume = "148",
pages = "112--20",
journal = "Advances in Biophysical Chemistry",
issn = "1057-8943",
publisher = "J A I Press Inc.",
number = "1-3",

}

RIS

TY - JOUR

T1 - In vitro association of fragments of a beta-sheet membrane protein

AU - Debnath, D

AU - Nielsen, K L

AU - Otzen, D E

PY - 2010/5/1

Y1 - 2010/5/1

N2 - Although the beta-barrel membrane protein OmpA can be produced in a biologically active form in E. coli from co-expressed fragments, the fragments have not been demonstrated to associate in vitro. We have produced 3 complementary fragment pairs of OmpA which can associate to form a folded complex according to the SDS band-shift assay. We are able to convert 25-35% of the fragment populations to non-covalent but SDS-stable complexes. The periplasmic chaperone Skp effectively prevented this association. Two separately expressed and purified overlapping fragments of OmpA can form a protease-resistant complex that undergoes the characteristic band-shift upon heating. Our work demonstrates that although membrane insertion and folding of beta-barrel membrane proteins may be a cooperative process, the fragments can associate in vitro without any additional components. However, the low yield and slow folding rates indicate that partially unfolded or destabilized beta-sheet membrane proteins can potentially engage in many non-native interactions.

AB - Although the beta-barrel membrane protein OmpA can be produced in a biologically active form in E. coli from co-expressed fragments, the fragments have not been demonstrated to associate in vitro. We have produced 3 complementary fragment pairs of OmpA which can associate to form a folded complex according to the SDS band-shift assay. We are able to convert 25-35% of the fragment populations to non-covalent but SDS-stable complexes. The periplasmic chaperone Skp effectively prevented this association. Two separately expressed and purified overlapping fragments of OmpA can form a protease-resistant complex that undergoes the characteristic band-shift upon heating. Our work demonstrates that although membrane insertion and folding of beta-barrel membrane proteins may be a cooperative process, the fragments can associate in vitro without any additional components. However, the low yield and slow folding rates indicate that partially unfolded or destabilized beta-sheet membrane proteins can potentially engage in many non-native interactions.

KW - Amino Acid Sequence

KW - Bacterial Outer Membrane Proteins

KW - DNA-Binding Proteins

KW - Escherichia coli Proteins

KW - Hot Temperature

KW - Kinetics

KW - Models, Molecular

KW - Molecular Chaperones

KW - Peptide Hydrolases

KW - Protein Folding

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - Substrate Specificity

U2 - 10.1016/j.bpc.2010.03.004

DO - 10.1016/j.bpc.2010.03.004

M3 - Journal article

C2 - 20356666

VL - 148

SP - 112

EP - 120

JO - Advances in Biophysical Chemistry

JF - Advances in Biophysical Chemistry

SN - 1057-8943

IS - 1-3

ER -