Facile backbone structure determination of human membrane proteins by NMR spectroscopy

Christian Klammt; Innokentiy Maslennikov; Monika Bayrhuber; Cédric Eichmann; Navratna Vajpai; Ellis Jeremy Chua Chiu; Katherine Y. Blain; Luis Esquivies; June Hyun Jung Kwon; Bartosz Balana; Ursula Pieper; Andrej Sali; Paul A. Slesinger; Witek Kwiatkowski; Roland Riek; Senyon Choe

doi:10.1038/nmeth.2033

Facile backbone structure determination of human membrane proteins by NMR spectroscopy

Christian Klammt, Innokentiy Maslennikov, Monika Bayrhuber, Cédric Eichmann, Navratna Vajpai, Ellis Jeremy Chua Chiu, Katherine Y. Blain, Luis Esquivies, June Hyun Jung Kwon, Bartosz Balana, Ursula Pieper, Andrej Sali, Paul A. Slesinger, Witek Kwiatkowski, Roland Riek, Senyon Choe

Research output: Contribution to journal › Article › peer-review

81 Scopus citations

Abstract

Although nearly half of today's major pharmaceutical drugs target human integral membrane proteins (hIMPs), only 30 hIMP structures are currently available in the Protein Data Bank, largely owing to inefficiencies in protein production. Here we describe a strategy for the rapid structure determination of hIMPs, using solution NMR spectroscopy with systematically labeled proteins produced via cell-free expression. We report new backbone structures of six hIMPs, solved in only 18 months from 15 initial targets. Application of our protocols to an additional 135 hIMPs with molecular weight <30 kDa yielded 38 hIMPs suitable for structural characterization by solution NMR spectroscopy without additional optimization.

Original language	English
Pages (from-to)	834-839
Number of pages	6
Journal	Nature Methods
Volume	9
Issue number	8
DOIs	https://doi.org/10.1038/nmeth.2033
State	Published - Aug 2012
Externally published	Yes

Access to Document

10.1038/nmeth.2033

Cite this

Klammt, C., Maslennikov, I., Bayrhuber, M., Eichmann, C., Vajpai, N., Chiu, E. J. C., Blain, K. Y., Esquivies, L., Kwon, J. H. J., Balana, B., Pieper, U., Sali, A., Slesinger, P. A., Kwiatkowski, W., Riek, R., & Choe, S. (2012). Facile backbone structure determination of human membrane proteins by NMR spectroscopy. Nature Methods, 9(8), 834-839. https://doi.org/10.1038/nmeth.2033

@article{425c17801e674e6c8bda27dde59f0475,

title = "Facile backbone structure determination of human membrane proteins by NMR spectroscopy",

abstract = "Although nearly half of today's major pharmaceutical drugs target human integral membrane proteins (hIMPs), only 30 hIMP structures are currently available in the Protein Data Bank, largely owing to inefficiencies in protein production. Here we describe a strategy for the rapid structure determination of hIMPs, using solution NMR spectroscopy with systematically labeled proteins produced via cell-free expression. We report new backbone structures of six hIMPs, solved in only 18 months from 15 initial targets. Application of our protocols to an additional 135 hIMPs with molecular weight <30 kDa yielded 38 hIMPs suitable for structural characterization by solution NMR spectroscopy without additional optimization.",

author = "Christian Klammt and Innokentiy Maslennikov and Monika Bayrhuber and C{\'e}dric Eichmann and Navratna Vajpai and Chiu, {Ellis Jeremy Chua} and Blain, {Katherine Y.} and Luis Esquivies and Kwon, {June Hyun Jung} and Bartosz Balana and Ursula Pieper and Andrej Sali and Slesinger, {Paul A.} and Witek Kwiatkowski and Roland Riek and Senyon Choe",

note = "Funding Information: We thank G. Louie for comments in preparation of the manuscript, A.S. Arseniev for suggestions on the spin-labeling procedure and S. Maslennikov for writing the atomDistancer program. C.K. thanks the Pioneer Foundation for a Pioneer Fund Postdoctoral Scholar Award. This work has been partly supported by US National Institutes of Health (S.C.: GM098630, GM095623; A.S. and U.P.: GM094662, GM094625 FDP, and GM54762), Incheon Free Economic Zone and the World Class University Program (Korea).",

year = "2012",

month = aug,

doi = "10.1038/nmeth.2033",

language = "English",

volume = "9",

pages = "834--839",

journal = "Nature Methods",

issn = "1548-7091",

publisher = "Nature Publishing Group",

number = "8",

}

TY - JOUR

T1 - Facile backbone structure determination of human membrane proteins by NMR spectroscopy

AU - Klammt, Christian

AU - Maslennikov, Innokentiy

AU - Bayrhuber, Monika

AU - Eichmann, Cédric

AU - Vajpai, Navratna

AU - Chiu, Ellis Jeremy Chua

AU - Blain, Katherine Y.

AU - Esquivies, Luis

AU - Kwon, June Hyun Jung

AU - Balana, Bartosz

AU - Pieper, Ursula

AU - Sali, Andrej

AU - Slesinger, Paul A.

AU - Kwiatkowski, Witek

AU - Riek, Roland

AU - Choe, Senyon

N1 - Funding Information: We thank G. Louie for comments in preparation of the manuscript, A.S. Arseniev for suggestions on the spin-labeling procedure and S. Maslennikov for writing the atomDistancer program. C.K. thanks the Pioneer Foundation for a Pioneer Fund Postdoctoral Scholar Award. This work has been partly supported by US National Institutes of Health (S.C.: GM098630, GM095623; A.S. and U.P.: GM094662, GM094625 FDP, and GM54762), Incheon Free Economic Zone and the World Class University Program (Korea).

PY - 2012/8

Y1 - 2012/8

N2 - Although nearly half of today's major pharmaceutical drugs target human integral membrane proteins (hIMPs), only 30 hIMP structures are currently available in the Protein Data Bank, largely owing to inefficiencies in protein production. Here we describe a strategy for the rapid structure determination of hIMPs, using solution NMR spectroscopy with systematically labeled proteins produced via cell-free expression. We report new backbone structures of six hIMPs, solved in only 18 months from 15 initial targets. Application of our protocols to an additional 135 hIMPs with molecular weight <30 kDa yielded 38 hIMPs suitable for structural characterization by solution NMR spectroscopy without additional optimization.

AB - Although nearly half of today's major pharmaceutical drugs target human integral membrane proteins (hIMPs), only 30 hIMP structures are currently available in the Protein Data Bank, largely owing to inefficiencies in protein production. Here we describe a strategy for the rapid structure determination of hIMPs, using solution NMR spectroscopy with systematically labeled proteins produced via cell-free expression. We report new backbone structures of six hIMPs, solved in only 18 months from 15 initial targets. Application of our protocols to an additional 135 hIMPs with molecular weight <30 kDa yielded 38 hIMPs suitable for structural characterization by solution NMR spectroscopy without additional optimization.

UR - http://www.scopus.com/inward/record.url?scp=84864426351&partnerID=8YFLogxK

U2 - 10.1038/nmeth.2033

DO - 10.1038/nmeth.2033

M3 - Article

C2 - 22609626

AN - SCOPUS:84864426351

SN - 1548-7091

VL - 9

SP - 834

EP - 839

JO - Nature Methods

JF - Nature Methods

IS - 8

ER -

Facile backbone structure determination of human membrane proteins by NMR spectroscopy

Abstract

Access to Document

Fingerprint

Cite this