ABCMdb

PMID: 19733108

Grant CV, Yang Y, Glibowicka M, Wu CH, Park SH, Deber CM, Opella SJ
A Modified Alderman-Grant Coil makes possible an efficient cross-coil probe for high field solid-state NMR of lossy biological samples.
J Magn Reson. 2009 Nov;201(1):87-92. Epub 2009 Aug 15., [PubMed]

Sentences

No. Mutations Sentence Comment

94 ABCC7 p.Val232Asp The initial samples containing the TM3/4 V232D segment of CFTR would form ordered and well-aligned bicelles only at low concentrations, near the threshold of solid-state NMR detection. Login to comment 111 ABCC7 p.Val232Asp (A) One-dimensional 15 N chemical shift spectrum of uniformly 15 N-labeled CFTR TM3/4 V232D. Login to comment 124 ABCC7 p.Val232Asp Expression and purification The 15 N labeled form of the CFTR TM3/4 V232D helical hairpin construct was expressed and purified as previously described [26-28]. Login to comment 130 ABCC7 p.Val232Asp Prior to cell growth, the medium was supplemented with biotin and thiamine (1 mg/L of each); sterile MgSO4 and CaCl2 stock solutions to final concentrations of 1 mM and 0.3 mM, respectively; 3 g of glucose for expression of 15 N isotopically labeled TM3/4 V232D. Login to comment 136 ABCC7 p.Val232Asp Thrombin-treated TM3/4 V232D was purified by RP-HPLC on a C4 semipreparative column (Phenomenex) using an acetonitrile gradient. Login to comment 140 ABCC7 p.Val232Asp 18 mg of TM3/4 V232D (>95% pure) per 1 L of minimal M9 medium. Login to comment 144 ABCC7 p.Val232Asp Bicelle sample preparation The uniformly 15 N-labeled TM3/4 V232D segment of CFTR and Pf1 major coat protein were reconstituted into bicelles as previously described [31,32]. Login to comment