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Expression of doubly labeled Saccharomyces cerevisiae iso ‐1 ferricytochrome c and 1 H, 13 C and 15 N chemical shift assignments by multidimensional NMR
Author(s) -
Szabo Christina M,
Sanders Lori K,
Le H.Carl,
Chien Ellen Y.T,
Oldfield Eric
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)02032-9
Subject(s) - chemical shift , chemistry , saccharomyces cerevisiae , amide , stereochemistry , crystallography , analytical chemistry (journal) , yeast , biochemistry , organic chemistry
We have expressed [U‐ 13 C, 15 N]‐labeled Saccharomyces cerevisiae iso ‐1 cytochrome c C102T;K72A in Escherichia coli with a yield of 11 mg/l of growth medium. Nuclear magnetic resonance (NMR) studies were conducted on the Fe 3+ form of the protein. We report herein chemical shift assignments for amide 1 H and 15 N, 13 C°, 13 C α , 13 C β , 1 H α and 1 H β resonances based upon a series of three‐dimensional NMR experiments: HNCA, HN(CO)CA, HNCO, HN(CA)CO, HNCACB, HCA(CO)N, HCCH‐TOCSY and HBHA(CBCA)NH. An investigation of the chemical shifts of the threonine residues was also made by using density functional theory in order to help solve discrepancies between 15 N chemical shift assignments reported in this study and those reported previously.