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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.

SUBMITTER: Klammt C

PROVIDER: S-EPMC3723349 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature

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Facile backbone structure determination of human membrane proteins by NMR spectroscopy.

Klammt Christian C Maslennikov Innokentiy I Bayrhuber Monika M Eichmann Cédric C Vajpai Navratna N Chiu Ellis Jeremy Chua EJ Blain Katherine Y KY Esquivies Luis L Kwon June Hyun Jung JH Balana Bartosz B Pieper Ursula U Sali Andrej A Slesinger Paul A PA Kwiatkowski Witek W Riek Roland R Choe Senyon S

Nature methods 20120520 8

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 in ...[more]

PMID: 22609626

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