2jfe: Difference between revisions
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==Overview== | ==Overview== | ||
Human cytosolic beta-glucosidase (hCBG) is a xenobiotic-metabolizing, enzyme that hydrolyses certain flavonoid glucosides, with specificity, depending on the aglycone moiety, the type of sugar and the linkage, between them. In this study, the substrate preference of this enzyme was, investigated by mutational analysis, X-ray crystallography and homology, modelling. The crystal structure of hCBG was solved by the molecular, replacement method and refined at 2.7 A resolution. The main-chain fold of, the enzyme belongs to the (beta/alpha)(8) barrel structure, which is, common to family 1 glycoside hydrolases. The active site is located at the, bottom of a pocket (about 16 A deep) formed by large surface loops, surrounding the C termini of the barrel of beta-strands. As for all the, clan of GH-A enzymes, the two catalytic glutamate residues are located on, strand 4 (the acid/base Glu165) and on strand 7 (the nucleophile Glu373)., Although many features of hCBG were shown to be very similar to previously, described enzymes from this family, crucial differences were observed in, the surface loops surrounding the aglycone binding site, and these are, likely to strongly influence the substrate specificity. The positioning of, a substrate molecule (quercetin-4'-glucoside) by homology modelling, revealed that hydrophobic interactions dominate the binding of the, aglycone moiety. In particular, Val168, Trp345, Phe225, Phe179, Phe334 and, Phe433 were identified as likely to be important in determining substrate, specificity in hCBG, and site-directed mutagenesis supported a key role, for some of these residues. | Human cytosolic beta-glucosidase (hCBG) is a xenobiotic-metabolizing, enzyme that hydrolyses certain flavonoid glucosides, with specificity, depending on the aglycone moiety, the type of sugar and the linkage, between them. In this study, the substrate preference of this enzyme was, investigated by mutational analysis, X-ray crystallography and homology, modelling. The crystal structure of hCBG was solved by the molecular, replacement method and refined at 2.7 A resolution. The main-chain fold of, the enzyme belongs to the (beta/alpha)(8) barrel structure, which is, common to family 1 glycoside hydrolases. The active site is located at the, bottom of a pocket (about 16 A deep) formed by large surface loops, surrounding the C termini of the barrel of beta-strands. As for all the, clan of GH-A enzymes, the two catalytic glutamate residues are located on, strand 4 (the acid/base Glu165) and on strand 7 (the nucleophile Glu373)., Although many features of hCBG were shown to be very similar to previously, described enzymes from this family, crucial differences were observed in, the surface loops surrounding the aglycone binding site, and these are, likely to strongly influence the substrate specificity. The positioning of, a substrate molecule (quercetin-4'-glucoside) by homology modelling, revealed that hydrophobic interactions dominate the binding of the, aglycone moiety. In particular, Val168, Trp345, Phe225, Phe179, Phe334 and, Phe433 were identified as likely to be important in determining substrate, specificity in hCBG, and site-directed mutagenesis supported a key role, for some of these residues. | ||
==Disease== | |||
Known disease associated with this structure: Corticosteroid-binding globulin deficiency OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=122500 122500]] | |||
==About this Structure== | ==About this Structure== | ||
2JFE is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Active as [http://en.wikipedia.org/wiki/Beta-glucosidase Beta-glucosidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.21 3.2.1.21] Known structural/functional Site: <scene name='pdbsite=AC1:Nag Binding Site For Chain X'>AC1</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2JFE OCA]. | 2JFE is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Active as [http://en.wikipedia.org/wiki/Beta-glucosidase Beta-glucosidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.21 3.2.1.21] Known structural/functional Site: <scene name='pdbsite=AC1:Nag+Binding+Site+For+Chain+X'>AC1</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2JFE OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: polymorphism]] | [[Category: polymorphism]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Feb 3 10:45:08 2008'' | ||
Revision as of 11:45, 3 February 2008
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THE CRYSTAL STRUCTURE OF HUMAN CYTOSOLIC BETA-GLUCOSIDASE
OverviewOverview
Human cytosolic beta-glucosidase (hCBG) is a xenobiotic-metabolizing, enzyme that hydrolyses certain flavonoid glucosides, with specificity, depending on the aglycone moiety, the type of sugar and the linkage, between them. In this study, the substrate preference of this enzyme was, investigated by mutational analysis, X-ray crystallography and homology, modelling. The crystal structure of hCBG was solved by the molecular, replacement method and refined at 2.7 A resolution. The main-chain fold of, the enzyme belongs to the (beta/alpha)(8) barrel structure, which is, common to family 1 glycoside hydrolases. The active site is located at the, bottom of a pocket (about 16 A deep) formed by large surface loops, surrounding the C termini of the barrel of beta-strands. As for all the, clan of GH-A enzymes, the two catalytic glutamate residues are located on, strand 4 (the acid/base Glu165) and on strand 7 (the nucleophile Glu373)., Although many features of hCBG were shown to be very similar to previously, described enzymes from this family, crucial differences were observed in, the surface loops surrounding the aglycone binding site, and these are, likely to strongly influence the substrate specificity. The positioning of, a substrate molecule (quercetin-4'-glucoside) by homology modelling, revealed that hydrophobic interactions dominate the binding of the, aglycone moiety. In particular, Val168, Trp345, Phe225, Phe179, Phe334 and, Phe433 were identified as likely to be important in determining substrate, specificity in hCBG, and site-directed mutagenesis supported a key role, for some of these residues.
DiseaseDisease
Known disease associated with this structure: Corticosteroid-binding globulin deficiency OMIM:[122500]
About this StructureAbout this Structure
2JFE is a Single protein structure of sequence from Homo sapiens. Active as Beta-glucosidase, with EC number 3.2.1.21 Known structural/functional Site: . Full crystallographic information is available from OCA.
ReferenceReference
The Crystal Structure of Human Cytosolic beta-Glucosidase Unravels the Substrate Aglycone Specificity of a Family 1 Glycoside Hydrolase., Tribolo S, Berrin JG, Kroon PA, Czjzek M, Juge N, J Mol Biol. 2007 Jul 27;370(5):964-75. Epub 2007 May 18. PMID:17555766
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