|
|
| (17 intermediate revisions not shown.) |
| Line 1: |
Line 1: |
| - | [[Image:1sua.gif|left|200px]]<br /><applet load="1sua" size="450" color="white" frame="true" align="right" spinBox="true" | |
| - | caption="1sua, resolution 2.1Å" /> | |
| - | '''SUBTILISIN BPN''''<br /> | |
| | | | |
| - | ==Overview== | + | ==SUBTILISIN BPN'== |
| - | The three-dimensional structure of a subtilisin BPN' construct that was, produced and folded without its prodomain shows the tertiary structure is, nearly identical to the wild-type enzyme and not a folding intermediate., The subtilisin BPN' variant, Sbt70, was cloned and expressed in, Escherichia coli without the prodomain, the 77-residue N-terminal domain, that catalyzes the folding of the enzyme into its native tertiary, structure. Sbt70 has the high-affinity calcium-binding loop, residues 75, to 83, deleted. Such calcium-independent forms of subtilisin BPN' refold, independently while retaining high levels of activity [Bryan et al., Biochemistry, 31:4937-4945, 1992]. Sbt70 has, in addition, seven, stabilizing mutations, K43N, M50F, A73L, Q206V, Y217K, N218S, Q271E, and, the active site serine has been replaced with alanine to prevent, autolysis. The purified Sbt70 folded spontaneously without the prodomain, and crystallized at room temperature. Crystals of Sbt70 belong to space, group P2(1)2(1)2(1) with unit cell parameters a = 53.5 A, b = 60.3 A, and, c = 83.4 A. Comparison of the refined structure with other high-resolution, structures of subtilisin BPN' establishes that the conformation of Sbt70, is essentially the same as that previously determined for other, calcium-independent forms and that of other wild-type subtilisin BPN', structures, all folded in the presence of the prodomain. These findings, confirm the results of previous solution studies that showed subtilisin, BPN' can be refolded into a native conformation without the presence of, the prodomain [Bryan et al., Biochemistry 31:4937-4945, 1992]. The, structure analysis also provides the first descriptions of four, stabilizing mutations, K43N, A73L, Q206V, and Q271E, and provides details, of the interaction between the enzyme and the Ala-Leu-Ala-Leu tetrapeptide, found in the active-site cleft. | + | <StructureSection load='1sua' size='340' side='right'caption='[[1sua]], [[Resolution|resolution]] 2.10Å' scene=''> |
| | + | == Structural highlights == |
| | + | <table><tr><td colspan='2'>[[1sua]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1SUA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1SUA FirstGlance]. <br> |
| | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.1Å</td></tr> |
| | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1sua FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1sua OCA], [https://pdbe.org/1sua PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1sua RCSB], [https://www.ebi.ac.uk/pdbsum/1sua PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1sua ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/SUBT_BACAM SUBT_BACAM] Subtilisin is an extracellular alkaline serine protease, it catalyzes the hydrolysis of proteins and peptide amides. Has a high substrate specificity to fibrin.<ref>PMID:12524032</ref> |
| | + | == Evolutionary Conservation == |
| | + | [[Image:Consurf_key_small.gif|200px|right]] |
| | + | Check<jmol> |
| | + | <jmolCheckbox> |
| | + | <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/su/1sua_consurf.spt"</scriptWhenChecked> |
| | + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked> |
| | + | <text>to colour the structure by Evolutionary Conservation</text> |
| | + | </jmolCheckbox> |
| | + | </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=1sua ConSurf]. |
| | + | <div style="clear:both"></div> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | The three-dimensional structure of a subtilisin BPN' construct that was produced and folded without its prodomain shows the tertiary structure is nearly identical to the wild-type enzyme and not a folding intermediate. The subtilisin BPN' variant, Sbt70, was cloned and expressed in Escherichia coli without the prodomain, the 77-residue N-terminal domain that catalyzes the folding of the enzyme into its native tertiary structure. Sbt70 has the high-affinity calcium-binding loop, residues 75 to 83, deleted. Such calcium-independent forms of subtilisin BPN' refold independently while retaining high levels of activity [Bryan et al., Biochemistry, 31:4937-4945, 1992]. Sbt70 has, in addition, seven stabilizing mutations, K43N, M50F, A73L, Q206V, Y217K, N218S, Q271E, and the active site serine has been replaced with alanine to prevent autolysis. The purified Sbt70 folded spontaneously without the prodomain and crystallized at room temperature. Crystals of Sbt70 belong to space group P2(1)2(1)2(1) with unit cell parameters a = 53.5 A, b = 60.3 A, and c = 83.4 A. Comparison of the refined structure with other high-resolution structures of subtilisin BPN' establishes that the conformation of Sbt70 is essentially the same as that previously determined for other calcium-independent forms and that of other wild-type subtilisin BPN' structures, all folded in the presence of the prodomain. These findings confirm the results of previous solution studies that showed subtilisin BPN' can be refolded into a native conformation without the presence of the prodomain [Bryan et al., Biochemistry 31:4937-4945, 1992]. The structure analysis also provides the first descriptions of four stabilizing mutations, K43N, A73L, Q206V, and Q271E, and provides details of the interaction between the enzyme and the Ala-Leu-Ala-Leu tetrapeptide found in the active-site cleft. |
| | | | |
| - | ==About this Structure==
| + | Crystal structure of calcium-independent subtilisin BPN' with restored thermal stability folded without the prodomain.,Almog O, Gallagher T, Tordova M, Hoskins J, Bryan P, Gilliland GL Proteins. 1998 Apr 1;31(1):21-32. PMID:9552156<ref>PMID:9552156</ref> |
| - | 1SUA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1SUA OCA].
| + | |
| | | | |
| - | ==Reference==
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| - | Crystal structure of calcium-independent subtilisin BPN' with restored thermal stability folded without the prodomain., Almog O, Gallagher T, Tordova M, Hoskins J, Bryan P, Gilliland GL, Proteins. 1998 Apr 1;31(1):21-32. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=9552156 9552156]
| + | </div> |
| - | [[Category: Bacillus amyloliquefaciens]]
| + | <div class="pdbe-citations 1sua" style="background-color:#fffaf0;"></div> |
| - | [[Category: Single protein]]
| + | |
| - | [[Category: Subtilisin]]
| + | |
| - | [[Category: Almog, O.]]
| + | |
| - | [[Category: Gilliland, G.L.]]
| + | |
| - | [[Category: complex (hydrolase/peptide)]]
| + | |
| - | [[Category: hydrolase]]
| + | |
| - | [[Category: serine proteinase]]
| + | |
| | | | |
| - | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 02:40:28 2007''
| + | ==See Also== |
| | + | *[[Subtilisin 3D structures|Subtilisin 3D structures]] |
| | + | == References == |
| | + | <references/> |
| | + | __TOC__ |
| | + | </StructureSection> |
| | + | [[Category: Bacillus amyloliquefaciens]] |
| | + | [[Category: Large Structures]] |
| | + | [[Category: Almog O]] |
| | + | [[Category: Gilliland GL]] |
| Structural highlights
Function
SUBT_BACAM Subtilisin is an extracellular alkaline serine protease, it catalyzes the hydrolysis of proteins and peptide amides. Has a high substrate specificity to fibrin.[1]
Evolutionary Conservation
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
The three-dimensional structure of a subtilisin BPN' construct that was produced and folded without its prodomain shows the tertiary structure is nearly identical to the wild-type enzyme and not a folding intermediate. The subtilisin BPN' variant, Sbt70, was cloned and expressed in Escherichia coli without the prodomain, the 77-residue N-terminal domain that catalyzes the folding of the enzyme into its native tertiary structure. Sbt70 has the high-affinity calcium-binding loop, residues 75 to 83, deleted. Such calcium-independent forms of subtilisin BPN' refold independently while retaining high levels of activity [Bryan et al., Biochemistry, 31:4937-4945, 1992]. Sbt70 has, in addition, seven stabilizing mutations, K43N, M50F, A73L, Q206V, Y217K, N218S, Q271E, and the active site serine has been replaced with alanine to prevent autolysis. The purified Sbt70 folded spontaneously without the prodomain and crystallized at room temperature. Crystals of Sbt70 belong to space group P2(1)2(1)2(1) with unit cell parameters a = 53.5 A, b = 60.3 A, and c = 83.4 A. Comparison of the refined structure with other high-resolution structures of subtilisin BPN' establishes that the conformation of Sbt70 is essentially the same as that previously determined for other calcium-independent forms and that of other wild-type subtilisin BPN' structures, all folded in the presence of the prodomain. These findings confirm the results of previous solution studies that showed subtilisin BPN' can be refolded into a native conformation without the presence of the prodomain [Bryan et al., Biochemistry 31:4937-4945, 1992]. The structure analysis also provides the first descriptions of four stabilizing mutations, K43N, A73L, Q206V, and Q271E, and provides details of the interaction between the enzyme and the Ala-Leu-Ala-Leu tetrapeptide found in the active-site cleft.
Crystal structure of calcium-independent subtilisin BPN' with restored thermal stability folded without the prodomain.,Almog O, Gallagher T, Tordova M, Hoskins J, Bryan P, Gilliland GL Proteins. 1998 Apr 1;31(1):21-32. PMID:9552156[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Peng Y, Huang Q, Zhang RH, Zhang YZ. Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food. Comp Biochem Physiol B Biochem Mol Biol. 2003 Jan;134(1):45-52. PMID:12524032
- ↑ Almog O, Gallagher T, Tordova M, Hoskins J, Bryan P, Gilliland GL. Crystal structure of calcium-independent subtilisin BPN' with restored thermal stability folded without the prodomain. Proteins. 1998 Apr 1;31(1):21-32. PMID:9552156
|
