嗜热古菌原Tk-枯草杆菌蛋白酶活性位点突变体的结晶及初步X射线衍射研究
Crystallization and preliminary X-ray diffraction study of an active-site mutant of pro-Tk-subtilisin from a hyperthermophilic archaeon.
作者信息
Tanaka Shun-ichi, Saito Kenji, Chon Hyongi, Matsumura Hiroyoshi, Koga Yuichi, Takano Kazufumi, Kanaya Shigenori
机构信息
Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Sep 1;62(Pt 9):902-5. doi: 10.1107/S1744309106030454. Epub 2006 Aug 18.
Abstract
Crystallization of and preliminary crystallographic studies on an active-site mutant of pro-Tk-subtilisin from the hyperthermophilic archaeon Thermococcus kodakaraensis were performed. The crystal was grown at 277 K by the sitting-drop vapour-diffusion method. Native X-ray diffraction data were collected to 2.3 A resolution using synchrotron radiation from station BL41XU at SPring-8. The crystal belongs to the orthorhombic space group I222, with unit-cell parameters a = 92.69, b = 121.78, c = 77.53 A. Assuming the presence of one molecule per asymmetric unit, the Matthews coefficient V(M) was calculated to be 2.6 A(3) Da(-1) and the solvent content was 53.1%.
摘要
对嗜热古菌柯达嗜热栖热菌（Thermococcus kodakaraensis）的前体枯草杆菌蛋白酶（pro-Tk-subtilisin）活性位点突变体进行了结晶及初步晶体学研究。通过坐滴气相扩散法在277 K下生长晶体。使用SPring-8的BL41XU站的同步辐射收集了分辨率为2.3 Å的天然X射线衍射数据。该晶体属于正交晶系空间群I222，晶胞参数a = 92.69，b = 121.78，c = 77.53 Å。假设每个不对称单元存在一个分子，计算得到的马修斯系数V(M)为2.6 ų Da⁻¹，溶剂含量为53.1%。
相似文献
1
Crystallization and preliminary X-ray diffraction study of an active-site mutant of pro-Tk-subtilisin from a hyperthermophilic archaeon.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Sep 1;62(Pt 9):902-5. doi: 10.1107/S1744309106030454. Epub 2006 Aug 18.
2
Crystallization and preliminary X-ray diffraction study of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Feb 1;63(Pt 2):126-9. doi: 10.1107/S1744309107001388. Epub 2007 Jan 27.
3
Crystal structure of Tk-subtilisin folded without propeptide: requirement of propeptide for acceleration of folding.
FEBS Lett. 2008 Nov 26;582(28):3875-8. doi: 10.1016/j.febslet.2008.10.025. Epub 2008 Oct 23.
4
Increase in activation rate of Pro-Tk-subtilisin by a single nonpolar-to-polar amino acid substitution at the hydrophobic core of the propeptide domain.
Protein Sci. 2013 Dec;22(12):1711-21. doi: 10.1002/pro.2371. Epub 2013 Oct 19.
5
Requirement of insertion sequence IS1 for thermal adaptation of Pro-Tk-subtilisin from hyperthermophilic archaeon.
Extremophiles. 2012 Nov;16(6):841-51. doi: 10.1007/s00792-012-0479-3. Epub 2012 Sep 21.
6
Crystallization and preliminary crystallographic analysis of type 1 RNase H from the hyperthermophilic archaeon Sulfolobus tokodaii 7.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Aug 1;62(Pt 8):781-4. doi: 10.1107/S1744309106024420. Epub 2006 Jul 25.
7
Crystallization and preliminary crystallographic studies of a ribosomal protein L30e from the hyperthermophilic archaeon Thermococcus celer.
Acta Crystallogr D Biol Crystallogr. 2001 Jun;57(Pt 6):865-6. doi: 10.1107/s0907444901004607. Epub 2001 May 25.
8
Crystallization and preliminary X-ray analysis of TBP-interacting protein from the hyperthermophilic archaeon Thermococcus kodakaraensis strain KOD1.
Acta Crystallogr D Biol Crystallogr. 2003 Feb;59(Pt 2):372-4. doi: 10.1107/s090744490202142x. Epub 2003 Jan 23.
9
Crystallization and preliminary X-ray diffraction analysis of L-threonine dehydrogenase (TDH) from the hyperthermophilic archaeon Thermococcus kodakaraensis.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Sep 1;64(Pt 9):828-30. doi: 10.1107/S1744309108025384. Epub 2008 Aug 20.
10
Crystallization and preliminary X-ray diffraction analysis of L-aminoacylase from the hyperthermophilic archaeon Thermococcus litoralis.
Acta Crystallogr D Biol Crystallogr. 2002 Mar;58(Pt 3):507-10. doi: 10.1107/s0907444901021266. Epub 2002 Feb 21.
引用本文的文献
1
Identification, overexpression, purification, and biochemical characterization of a novel hyperthermostable keratinase from .
3 Biotech. 2021 Jan;11(1):2. doi: 10.1007/s13205-020-02538-1. Epub 2020 Nov 27.
2
Increase in activation rate of Pro-Tk-subtilisin by a single nonpolar-to-polar amino acid substitution at the hydrophobic core of the propeptide domain.
Protein Sci. 2013 Dec;22(12):1711-21. doi: 10.1002/pro.2371. Epub 2013 Oct 19.
3
Enzymatic activity of a subtilisin homolog, Tk-SP, from Thermococcus kodakarensis in detergents and its ability to degrade the abnormal prion protein.
BMC Biotechnol. 2013 Feb 28;13:19. doi: 10.1186/1472-6750-13-19.
本文引用的文献
1
Processing of X-ray diffraction data collected in oscillation mode.
Methods Enzymol. 1997;276:307-26. doi: 10.1016/S0076-6879(97)76066-X.
2
Ca2+-dependent maturation of subtilisin from a hyperthermophilic archaeon, Thermococcus kodakaraensis: the propeptide is a potent inhibitor of the mature domain but is not required for its folding.
Appl Environ Microbiol. 2006 Jun;72(6):4154-62. doi: 10.1128/AEM.02696-05.
3
Folding pathway mediated by an intramolecular chaperone: intrinsically unstructured propeptide modulates stochastic activation of subtilisin.
J Mol Biol. 2005 Mar 25;347(2):367-83. doi: 10.1016/j.jmb.2005.01.028. Epub 2005 Jan 27.
4
Synthetic peptides derived from the prosegments of proprotein convertase 1/3 and furin are potent inhibitors of both enzymes.
Biochem J. 2003 Jul 1;373(Pt 1):231-9. doi: 10.1042/BJ20030120.
5
Folding pathway mediated by an intramolecular chaperone: dissecting conformational changes coincident with autoprocessing and the role of Ca(2+) in subtilisin maturation.
J Biochem. 2002 Jan;131(1):31-7. doi: 10.1093/oxfordjournals.jbchem.a003074.
6
Folding pathway mediated by an intramolecular chaperone: propeptide release modulates activation precision of pro-subtilisin.
J Biol Chem. 2001 Nov 30;276(48):44427-34. doi: 10.1074/jbc.M107573200. Epub 2001 Sep 27.
7
Active subtilisin-like protease from a hyperthermophilic archaeon in a form with a putative prosequence.
Appl Environ Microbiol. 2001 Jun;67(6):2445-52. doi: 10.1128/AEM.67.6.2445-2452.2001.
8
Folding pathway mediated by an intramolecular chaperone: the structural and functional characterization of the aqualysin I propeptide.
J Mol Biol. 2001 Jan 5;305(1):151-65. doi: 10.1006/jmbi.2000.4233.
9
Calcium-mediated thermostability in the subtilisin superfamily: the crystal structure of Bacillus Ak.1 protease at 1.8 A resolution.
J Mol Biol. 1999 Dec 10;294(4):1027-40. doi: 10.1006/jmbi.1999.3291.
10
Intramolecular chaperone and inhibitor activities of a propeptide from a bacterial zinc aminopeptidase.
Biochem J. 1999 Jul 1;341 ( Pt 1)(Pt 1):25-31.
Zotero 插件