TY - JOUR
T1 - High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements
AU - Mine, Shouhei
AU - Ueda, Tadashi
AU - Hashimoto, Yoshio
AU - Tanaka, Yoshitsugu
AU - Imoto, Taiji
N1 - Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1999/4/2
Y1 - 1999/4/2
N2 - The non-enzymatic deamidation of Asn to Asp is known to occur in proteins and peptides and is accelerated by phosphate buffer [Tyler-Cross, R. and Schirch, V. (1991) J. Biol. Chem. 25, 22549-22556]. We attempted to identify the site in lysozyme where a phosphate ion binds by means of 1H-15N HSQC measurements of 15N-labeled lysozyme, which was successfully obtained using Pichia pastoris. As a result, we found that the phosphate ion was preferentially bound to Asn-103 in hen lysozyme. The method presented here may be useful for identifying the binding site of a protein with low molecular weight substances. Copyright (C) 1999 Federation of European Biochemical Societies.
AB - The non-enzymatic deamidation of Asn to Asp is known to occur in proteins and peptides and is accelerated by phosphate buffer [Tyler-Cross, R. and Schirch, V. (1991) J. Biol. Chem. 25, 22549-22556]. We attempted to identify the site in lysozyme where a phosphate ion binds by means of 1H-15N HSQC measurements of 15N-labeled lysozyme, which was successfully obtained using Pichia pastoris. As a result, we found that the phosphate ion was preferentially bound to Asn-103 in hen lysozyme. The method presented here may be useful for identifying the binding site of a protein with low molecular weight substances. Copyright (C) 1999 Federation of European Biochemical Societies.
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U2 - 10.1016/S0014-5793(99)00332-4
DO - 10.1016/S0014-5793(99)00332-4
M3 - Article
C2 - 10217404
AN - SCOPUS:0032991055
VL - 448
SP - 33
EP - 37
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 1
ER -