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1bsi

From Proteopedia

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[[Image:1bsi.gif|left|200px]]
 
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{{Structure
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==HUMAN PANCREATIC ALPHA-AMYLASE FROM PICHIA PASTORIS, GLYCOSYLATED PROTEIN==
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|PDB= 1bsi |SIZE=350|CAPTION= <scene name='initialview01'>1bsi</scene>, resolution 2.&Aring;
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<StructureSection load='1bsi' size='340' side='right'caption='[[1bsi]], [[Resolution|resolution]] 2.00&Aring;' scene=''>
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|SITE=
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== Structural highlights ==
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|LIGAND= <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PCA:PYROGLUTAMIC+ACID'>PCA</scene>
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<table><tr><td colspan='2'>[[1bsi]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1BSI OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1BSI FirstGlance]. <br>
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|ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Alpha-amylase Alpha-amylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.1 3.2.1.1] </span>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2&#8491;</td></tr>
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|GENE=
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<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PCA:PYROGLUTAMIC+ACID'>PCA</scene></td></tr>
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|DOMAIN=
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1bsi FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1bsi OCA], [https://pdbe.org/1bsi PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1bsi RCSB], [https://www.ebi.ac.uk/pdbsum/1bsi PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1bsi ProSAT]</span></td></tr>
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|RELATEDENTRY=
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</table>
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|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1bsi FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1bsi OCA], [http://www.ebi.ac.uk/pdbsum/1bsi PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1bsi RCSB]</span>
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== Function ==
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}}
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[https://www.uniprot.org/uniprot/AMYP_HUMAN AMYP_HUMAN]
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/bs/1bsi_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1bsi ConSurf].
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<div style="clear:both"></div>
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'''HUMAN PANCREATIC ALPHA-AMYLASE FROM PICHIA PASTORIS, GLYCOSYLATED PROTEIN'''
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==See Also==
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*[[Amylase 3D structures|Amylase 3D structures]]
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__TOC__
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==Overview==
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</StructureSection>
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Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile.
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==About this Structure==
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1BSI is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1BSI OCA].
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==Reference==
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Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris., Rydberg EH, Sidhu G, Vo HC, Hewitt J, Cote HC, Wang Y, Numao S, MacGillivray RT, Overall CM, Brayer GD, Withers SG, Protein Sci. 1999 Mar;8(3):635-43. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/10091666 10091666]
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[[Category: Alpha-amylase]]
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[[Category: Homo sapiens]]
[[Category: Homo sapiens]]
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[[Category: Single protein]]
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[[Category: Large Structures]]
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[[Category: Brayer, G D.]]
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[[Category: Brayer GD]]
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[[Category: Cote, H C.F.]]
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[[Category: Cote HCF]]
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[[Category: Hewitt, J.]]
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[[Category: Hewitt J]]
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[[Category: Macgillivray, R T.A.]]
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[[Category: Macgillivray RTA]]
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[[Category: Numao, S.]]
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[[Category: Numao S]]
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[[Category: Overall, C M.]]
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[[Category: Overall CM]]
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[[Category: Rydberg, E H.]]
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[[Category: Rydberg EH]]
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[[Category: Sidhu, G.]]
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[[Category: Sidhu G]]
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[[Category: Vo, H C.]]
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[[Category: Vo HC]]
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[[Category: Wang, Y.]]
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[[Category: Wang Y]]
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[[Category: Withers, S G.]]
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[[Category: Withers SG]]
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[[Category: amylase]]
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[[Category: catalysis]]
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[[Category: diabetes]]
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[[Category: enzyme]]
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[[Category: glycosylated protein]]
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[[Category: human]]
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[[Category: hydrolase]]
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[[Category: mutagenesis]]
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[[Category: pancreatic]]
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[[Category: pichia pastori]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 19:07:22 2008''
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Current revision

HUMAN PANCREATIC ALPHA-AMYLASE FROM PICHIA PASTORIS, GLYCOSYLATED PROTEIN

PDB ID 1bsi

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