1nss: Difference between revisions
New page: left|200px<br /><applet load="1nss" size="450" color="white" frame="true" align="right" spinBox="true" caption="1nss, resolution 1.85Å" /> '''Crystal structure of... |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:1nss.gif|left|200px]]<br /><applet load="1nss" size=" | [[Image:1nss.gif|left|200px]]<br /><applet load="1nss" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1nss, resolution 1.85Å" /> | caption="1nss, resolution 1.85Å" /> | ||
'''Crystal structure of galactose mutarotase from Lactococcus lactis mutant D243A complexed with glucose'''<br /> | '''Crystal structure of galactose mutarotase from Lactococcus lactis mutant D243A complexed with glucose'''<br /> | ||
==Overview== | ==Overview== | ||
Galactose mutarotase catalyzes the first step in normal galactose | Galactose mutarotase catalyzes the first step in normal galactose metabolism by catalyzing the conversion of beta-D-galactose to alpha-D-galactose. The structure of the enzyme from Lactococcus lactis was recently solved in this laboratory and shown to be topologically similar to domain 5 of beta-galactosidase. From this initial X-ray analysis, four amino acid residues were demonstrated to be intimately involved in sugar binding to the protein: His 96, His 170, Asp 243, and Glu 304. Here we present a combined X-ray crystallographic and kinetic analysis designed to examine the role of these residues in the reaction mechanism of the enzyme. For this investigation, the following site-directed mutant proteins were prepared: H96N, H170N, D243N, D243A, E304Q, and E304A. All of the structures of these proteins, complexed with either glucose or galactose, were solved to a nominal resolution of 1.95 A or better, and their kinetic parameters were measured against D-galactose, D-glucose, L-arabinose, or D-xylose. From these studies, it can be concluded that Glu 304 and His 170 are critical for catalysis and that His 96 and Asp 243 are important for proper substrate positioning within the active site. Specifically, Glu 304 serves as the active site base to initiate the reaction by removing the proton from the C-1 hydroxyl group of the sugar substrate and His 170 functions as the active site acid to protonate the C-5 ring oxygen. | ||
==About this Structure== | ==About this Structure== | ||
1NSS is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactococcus_lactis Lactococcus lactis] with GLC and NA as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Aldose_1-epimerase Aldose 1-epimerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.3.3 5.1.3.3] Full crystallographic information is available from [http:// | 1NSS is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactococcus_lactis Lactococcus lactis] with <scene name='pdbligand=GLC:'>GLC</scene> and <scene name='pdbligand=NA:'>NA</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Aldose_1-epimerase Aldose 1-epimerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.3.3 5.1.3.3] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NSS OCA]. | ||
==Reference== | ==Reference== | ||
| Line 14: | Line 14: | ||
[[Category: Lactococcus lactis]] | [[Category: Lactococcus lactis]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Holden, H | [[Category: Holden, H M.]] | ||
[[Category: Thoden, J | [[Category: Thoden, J B.]] | ||
[[Category: GLC]] | [[Category: GLC]] | ||
[[Category: NA]] | [[Category: NA]] | ||
| Line 22: | Line 22: | ||
[[Category: mutarotase]] | [[Category: mutarotase]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:09:37 2008'' | ||
Revision as of 15:09, 21 February 2008
|
Crystal structure of galactose mutarotase from Lactococcus lactis mutant D243A complexed with glucose
OverviewOverview
Galactose mutarotase catalyzes the first step in normal galactose metabolism by catalyzing the conversion of beta-D-galactose to alpha-D-galactose. The structure of the enzyme from Lactococcus lactis was recently solved in this laboratory and shown to be topologically similar to domain 5 of beta-galactosidase. From this initial X-ray analysis, four amino acid residues were demonstrated to be intimately involved in sugar binding to the protein: His 96, His 170, Asp 243, and Glu 304. Here we present a combined X-ray crystallographic and kinetic analysis designed to examine the role of these residues in the reaction mechanism of the enzyme. For this investigation, the following site-directed mutant proteins were prepared: H96N, H170N, D243N, D243A, E304Q, and E304A. All of the structures of these proteins, complexed with either glucose or galactose, were solved to a nominal resolution of 1.95 A or better, and their kinetic parameters were measured against D-galactose, D-glucose, L-arabinose, or D-xylose. From these studies, it can be concluded that Glu 304 and His 170 are critical for catalysis and that His 96 and Asp 243 are important for proper substrate positioning within the active site. Specifically, Glu 304 serves as the active site base to initiate the reaction by removing the proton from the C-1 hydroxyl group of the sugar substrate and His 170 functions as the active site acid to protonate the C-5 ring oxygen.
About this StructureAbout this Structure
1NSS is a Single protein structure of sequence from Lactococcus lactis with and as ligands. Active as Aldose 1-epimerase, with EC number 5.1.3.3 Full crystallographic information is available from OCA.
ReferenceReference
The catalytic mechanism of galactose mutarotase., Thoden JB, Kim J, Raushel FM, Holden HM, Protein Sci. 2003 May;12(5):1051-9. PMID:12717027
Page seeded by OCA on Thu Feb 21 14:09:37 2008