DOI: 10.1110/ps.8.10.1990. Three‐dimensional structures of enzyme‐substrate complexes of the hydroxynitrile lyase from <i>hevea brasiliensis</i>

Three‐dimensional structures of enzyme‐substrate complexes of the hydroxynitrile lyase from hevea brasiliensis

10.1110/ps.8.10.1990

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractThe 3D structures of complexes between the hydroxynitrile lyase from Hevea brasiliensis (Hb‐HNL) and several substrate and/or inhibitor molecules, including trichloracetaldehyde, hexafluoracetone, acetone, and rhodanide, were determined by X‐ray crystallography. The complex with trichloracetaldehyde showed a covalent linkage between the protein and the inhibitor, which had apparently resulted from nucleophilic attack of the catalytic Ser80‐Oγ. All other complexes showed the substrate or inhibitor molecule merely hydrogen bonded to the protein. In addition, the native crystal structure of Hb‐HNL was redetermined at cryo‐temperature and at room temperature, eliminating previous uncertainties concerning residual electron density within the active site, and leading to the observation of two conserved water molecules. One of them was found to be conserved in all complex structures and appears to have mainly structural significance. The other water molecule is conserved in all structures except for the complex with rhodanide; it is hydrogen bonded to the imidazole of the catalytic His235 and appears to affect the Hb‐HNL catalyzed reaction. The observed 3D structural data suggest implications for the enzyme mechanism. It appears that the enzyme‐catalyzed cyanohydrin formation is unlikely to proceed via a hemiacetal or hemiketal intermediate covalently attached to the enzyme, despite the observation of such an intermediate for the complex with trichloracetaldehyde. Instead, the data are consistent with a mechanism where the incoming substrate is activated by hydrogen bonding with its carbonyl oxygen to the Ser80 and Thr11 hydroxy groups. A hydrogen cyanide molecule subsequently replaces a water molecule and is deprotonated presumably by the His235 base. Deprotonation is facilitated by the proximity of the positive charge of the Lys236 side chain.

Bibliography

Zuegg, J., Gruber, K., Gugganig, M., Wagner, U. G., & Kratky, C. (1999). Threeâdimensional structures of enzymeâsubstrate complexes of the hydroxynitrile lyase from hevea brasiliensis. Protein Science, 8(10), 1990â2000. Portico.

Authors 5

J. Zuegg (first)
K. Gruber (additional)
M. Gugganig (additional)
U. G. Wagner (additional)
C. Kratky (additional)

References 51 Referenced 67

{'key': 'e_1_2_1_2_1', 'first-page': '89', 'article-title': 'Classification of serine proteases derived from steric comparisons of their active site geometry, part II: Ser, His, Asp arrangements in proteolytic and nonproteolytic proteins', 'volume': '12', 'author': 'Barth A', 'year': '1994', 'journal-title': 'Drug Design Discovery'} / Drug Design Discovery / Classification of serine proteases derived from steric comparisons of their active site geometry, part II: Ser, His, Asp arrangements in proteolytic and nonproteolytic proteins by Barth A (1994)
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Dates

Type	When
Created	16 years, 6 months ago (Feb. 17, 2009, 4:35 p.m.)
Deposited	1 year, 10 months ago (Oct. 28, 2023, 1:03 p.m.)
Indexed	1 day, 12 hours ago (Sept. 2, 2025, 6:41 a.m.)
Issued	26 years, 8 months ago (Jan. 1, 1999)
Published	26 years, 8 months ago (Jan. 1, 1999)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	26 years, 8 months ago (Jan. 1, 1999)

Funders 0

None

BibTeX

@article{Zuegg_1999, title={Three‐dimensional structures of enzyme‐substrate complexes of the hydroxynitrile lyase from hevea brasiliensis}, volume={8}, ISSN={1469-896X}, url={http://dx.doi.org/10.1110/ps.8.10.1990}, DOI={10.1110/ps.8.10.1990}, number={10}, journal={Protein Science}, publisher={Wiley}, author={Zuegg, J. and Gruber, K. and Gugganig, M. and Wagner, U. G. and Kratky, C.}, year={1999}, month=jan, pages={1990–2000} }

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