Facile backbone structure determination of human membrane proteins by NMR spectroscopy
C Klammt, I Maslennikov, M Bayrhuber, C Eichmann… - Nature …, 2012 - nature.com
C Klammt, I Maslennikov, M Bayrhuber, C Eichmann, N Vajpai, EJC Chiu, KY Blain…
Nature methods, 2012•nature.comAlthough 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 …
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 …
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.
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