Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy

Katrine Østergaard Bugge; Helena Steinocher; Andrew J. Brooks; Kresten Lindorff-Larsen; Birthe Brandt Kragelund

doi:10.1021/acs.analchem.5b02365

Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy

Katrine Østergaard Bugge, Helena Steinocher, Andrew J. Brooks, Kresten Lindorff-Larsen, Birthe Brandt Kragelund

Biomolecular Sciences

6 Citationer (Scopus)

Abstract

Despite the biological and pharmaceutical significance of membrane proteins, their tertiary structures constitute less than 3% of known structures. One of the major obstacles for initiating structural studies of membrane proteins by NMR spectroscopy is the generation of high amounts of isotope-labeled protein. In this work, we have exploited the hydrophobic nature of membrane proteins to develop a simple and efficient production scheme for isotope-labeled single-pass transmembrane domains (TMDs) with or without intrinsically disordered regions. We have evaluated the applicability and limitations of the strategy using seven membrane protein variants that differ in their overall hydrophobicity and length and show a recovery for suitable variants of >70%. The developed production scheme is cost-efficient and easy to implement and has the potential to facilitate an increase in the number of structures of single-pass TMDs, which are difficult to solve by other means.

Originalsprog	Engelsk
Tidsskrift	Analytical Chemistry
Vol/bind	87
Udgave nummer	18
Sider (fra-til)	9126–9131
Antal sider	6
ISSN	0003-2700
DOI	https://doi.org/10.1021/acs.analchem.5b02365
Status	Udgivet - 15 sep. 2015

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10.1021/acs.analchem.5b02365

Citationsformater

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title = "Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy",

abstract = "Despite the biological and pharmaceutical significance of membrane proteins, their tertiary structures constitute less than 3% of known structures. One of the major obstacles for initiating structural studies of membrane proteins by NMR spectroscopy is the generation of high amounts of isotope-labeled protein. In this work, we have exploited the hydrophobic nature of membrane proteins to develop a simple and efficient production scheme for isotope-labeled single-pass transmembrane domains (TMDs) with or without intrinsically disordered regions. We have evaluated the applicability and limitations of the strategy using seven membrane protein variants that differ in their overall hydrophobicity and length and show a recovery for suitable variants of >70%. The developed production scheme is cost-efficient and easy to implement and has the potential to facilitate an increase in the number of structures of single-pass TMDs, which are difficult to solve by other means.",

author = "Bugge, {Katrine {\O}stergaard} and Helena Steinocher and Brooks, {Andrew J.} and Kresten Lindorff-Larsen and Kragelund, {Birthe Brandt}",

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T1 - Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy

AU - Bugge, Katrine Østergaard

AU - Steinocher, Helena

AU - Brooks, Andrew J.

AU - Lindorff-Larsen, Kresten

AU - Kragelund, Birthe Brandt

PY - 2015/9/15

Y1 - 2015/9/15

N2 - Despite the biological and pharmaceutical significance of membrane proteins, their tertiary structures constitute less than 3% of known structures. One of the major obstacles for initiating structural studies of membrane proteins by NMR spectroscopy is the generation of high amounts of isotope-labeled protein. In this work, we have exploited the hydrophobic nature of membrane proteins to develop a simple and efficient production scheme for isotope-labeled single-pass transmembrane domains (TMDs) with or without intrinsically disordered regions. We have evaluated the applicability and limitations of the strategy using seven membrane protein variants that differ in their overall hydrophobicity and length and show a recovery for suitable variants of >70%. The developed production scheme is cost-efficient and easy to implement and has the potential to facilitate an increase in the number of structures of single-pass TMDs, which are difficult to solve by other means.

AB - Despite the biological and pharmaceutical significance of membrane proteins, their tertiary structures constitute less than 3% of known structures. One of the major obstacles for initiating structural studies of membrane proteins by NMR spectroscopy is the generation of high amounts of isotope-labeled protein. In this work, we have exploited the hydrophobic nature of membrane proteins to develop a simple and efficient production scheme for isotope-labeled single-pass transmembrane domains (TMDs) with or without intrinsically disordered regions. We have evaluated the applicability and limitations of the strategy using seven membrane protein variants that differ in their overall hydrophobicity and length and show a recovery for suitable variants of >70%. The developed production scheme is cost-efficient and easy to implement and has the potential to facilitate an increase in the number of structures of single-pass TMDs, which are difficult to solve by other means.

U2 - 10.1021/acs.analchem.5b02365

DO - 10.1021/acs.analchem.5b02365

M3 - Journal article

C2 - 26309151

SN - 0003-2700

VL - 87

SP - 9126

EP - 9131

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 18

ER -

Exploiting hydrophobicity for efficient production of transmembrane helices for structure determination by NMR spectroscopy

Abstract

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