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1 H, 13 C, and 15 N resonance assignments of Fusarium solani pisi cutinase and preliminary features of the structure in solution
Author(s) -
Prompers Jeanine J.,
Hilbers Cornelis W.,
Groenewegen Anneke,
Schaik René C. Van,
Pepermans Henri A. M.
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560061111
Subject(s) - oxyanion , cutinase , oxyanion hole , heteronuclear molecule , chemistry , crystallography , stereochemistry , crystal structure , side chain , resonance (particle physics) , fusarium solani , nuclear magnetic resonance spectroscopy , hydrogen bond , molecule , enzyme , organic chemistry , physics , biology , particle physics , polymer , microbiology and biotechnology , catalysis
Essentially complete (96%) sequence‐specific assignments were made for the backbone and side‐chain 1 H, 13 C, and 15 N resonances of Fusarium solani pisi cutinase, produced as a 214‐residue heterologous protein in Escherichia coli , using heteronuclear NMR techniques. Three structural features were noticed during the assignment. (1) The secondary structure in solution corresponds mostly with the structure from X‐ray diffraction, suggesting that both structures are globally similar. (2) The H N of Ala 32 has a strongly upfield‐shifted resonance at 3.97 ppm, indicative of an amide‐aromatic hydrogen bond to the indole ring of Trp 69 that stabilizes the N‐terminal side of the parallel β‐sheet. (3) The NMR data suggest that the residues constituting the oxyanion hole are quite mobile in the free enzyme in solution, in contrast to the existence of a preformed oxyanion hole as observed in the crystal structure. Apparently, cutinase forms its oxyanion hole upon binding of the substrate like true lipases.