دورية أكاديمية
Hemiacetal stabilization in a chymotrypsin inhibitor complex and the reactivity of the hydroxyl group of the catalytic serine residue of chymotrypsin.
| العنوان: | Hemiacetal stabilization in a chymotrypsin inhibitor complex and the reactivity of the hydroxyl group of the catalytic serine residue of chymotrypsin. |
|---|---|
| المؤلفون: | Cleary JA; School of Biomolecular and Biomedical Science, Centre for Synthesis and Chemical Biology, Conway Institute, University College Dublin, Dublin 4, Ireland., Doherty W; School of Chemistry and Chemical Biology, Centre for Synthesis and Chemical Biology, Conway Institute, University College Dublin, Dublin 4, Ireland., Evans P; School of Chemistry and Chemical Biology, Centre for Synthesis and Chemical Biology, Conway Institute, University College Dublin, Dublin 4, Ireland., Malthouse JP; School of Biomolecular and Biomedical Science, Centre for Synthesis and Chemical Biology, Conway Institute, University College Dublin, Dublin 4, Ireland. Electronic address: J.Paul.G.Malthouse@ucd.ie. |
| المصدر: | Biochimica et biophysica acta [Biochim Biophys Acta] 2014 Jun; Vol. 1844 (6), pp. 1119-27. Date of Electronic Publication: 2014 Mar 21. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Pub. Co Country of Publication: Netherlands NLM ID: 0217513 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0006-3002 (Print) Linking ISSN: 00063002 NLM ISO Abbreviation: Biochim Biophys Acta Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam : Elsevier Pub. Co. |
| مواضيع طبية MeSH: | Chymotrypsin/*chemistry , Histidine/*chemistry , Oligopeptides/*chemistry , Protease Inhibitors/*chemistry , Serine/*chemistry, Animals ; Biocatalysis ; Catalytic Domain ; Cattle ; Glyoxal/chemistry ; Hydrogen Bonding ; Hydrogen-Ion Concentration ; Kinetics ; Nuclear Magnetic Resonance, Biomolecular ; Oligopeptides/chemical synthesis ; Protease Inhibitors/chemical synthesis ; Solutions ; Stereoisomerism ; Thermodynamics |
| مستخلص: | The aldehyde inhibitor Z-Ala-Ala-Phe-CHO has been synthesized and shown by (13)C-NMR to react with the active site serine hydroxyl group of alpha-chymotrypsin to form two diastereomeric hemiacetals. For both hemiacetals oxyanion formation occurs with a pKa value of ~7 showing that chymotrypsin reduces the oxyanion pKa values by ~5.6 pKa units and stabilizes the oxyanions of both diastereoisomers by ~32kJmol(-1). As pH has only a small effect on binding we conclude that oxyanion formation does not have a significant effect on binding the aldehyde inhibitor. By comparing the binding of Z-Ala-Ala-Phe-CHO with that of Z-Ala-Ala-Phe-H we estimate that the aldehyde group increases binding ~100 fold. At pH7.2 the effective molarity of the active site serine hydroxy group is ~6000 which is ~7× less effective than with the corresponding glyoxal inhibitor. Using (1)H-NMR we have shown that at both 4 and 25°C the histidine pKa is ~7.3 in free chymotrypsin and it is raised to ~8 when Z-Ala-Ala-Phe-CHO is bound. We conclude that oxyanion formation only has a minor role in raising the histidine pKa and that the aldehyde hydrogen must be replaced by a larger group to raise the histidine pKa>10 and give stereospecific formation of tetrahedral intermediates. The results show that a large increase in the pKa of the active site histidine is not needed for the active site serine hydroxyl group to have an effective molarity of 6000. (Copyright © 2014 Elsevier B.V. All rights reserved.) |
| References: | Biochemistry. 1987 May 5;26(9):2611-5. (PMID: 3607037) Biochemistry. 2001 Feb 27;40(8):2439-47. (PMID: 11327865) Biochem J. 1992 Sep 15;286 ( Pt 3):889-900. (PMID: 1417749) J Med Chem. 2010 Feb 25;53(4):1509-18. (PMID: 20112914) Biochemistry. 2007 Oct 2;46(39):11205-15. (PMID: 17824620) J Mol Biol. 1985 May 5;183(1):89-103. (PMID: 3892018) Biochemistry. 1973 Jan 2;12(1):47-51. (PMID: 4734224) Biochem J. 2002 Mar 1;362(Pt 2):339-47. (PMID: 11853541) Biochim Biophys Acta. 2009 Aug;1794(8):1251-8. (PMID: 19393346) Proc Natl Acad Sci U S A. 1985 Dec;82(23):7948-51. (PMID: 3934665) Biochem J. 1989 Mar 15;258(3):853-9. (PMID: 2730570) Biochemistry. 2009 Apr 21;48(15):3490-6. (PMID: 19281249) J Med Chem. 2011 Dec 8;54(23):7962-73. (PMID: 22014094) J Mol Biol. 1971 Sep 14;60(2):279-90. (PMID: 5099294) Biochemistry. 1979 Mar 6;18(5):921-6. (PMID: 420824) Biochemistry. 1979 Jan 23;18(2):349-56. (PMID: 33696) Biochemistry. 1987 Dec 1;26(24):7603-8. (PMID: 3427096) Eur J Biochem. 1980 Apr;105(3):581-5. (PMID: 6245886) J Biol Chem. 2007 Mar 16;282(11):7852-61. (PMID: 17213185) Biochemistry. 1996 Aug 27;35(34):11092-7. (PMID: 8780512) Arch Biochem Biophys. 1987 Feb 1;252(2):626-34. (PMID: 3813553) Biochemistry. 2012 Aug 7;51(31):6164-70. (PMID: 22757750) Biochemistry. 1991 Oct 15;30(41):10026-34. (PMID: 1911768) J Mol Biol. 1974 Jul 5;86(3):519-40. (PMID: 4852269) J Biol Chem. 1972 Dec 25;247(24):8195-7. (PMID: 4640942) Biochemistry. 1982 Sep 14;21(19):4679-86. (PMID: 7138821) J Mol Biol. 1972 Nov 14;71(2):507-11. (PMID: 4635995) Protein Sci. 1996 Apr;5(4):752-8. (PMID: 8845765) |
| معلومات مُعتمدة: | United Kingdom Wellcome Trust; 055637/Z/98 United Kingdom WT_ Wellcome Trust |
| فهرسة مساهمة: | Keywords: Aldehyde inhibitor; Chymotrypsin; Effective molarity; Hemiacetal; Oxyanion |
| المشرفين على المادة: | 0 (Oligopeptides) 0 (Protease Inhibitors) 0 (Solutions) 452VLY9402 (Serine) 4QD397987E (Histidine) 50NP6JJ975 (Glyoxal) EC 3.4.21.1 (Chymotrypsin) EC 3.4.21.1 (alpha-chymotrypsin) |
| تواريخ الأحداث: | Date Created: 20140325 Date Completed: 20140626 Latest Revision: 20220216 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC7185751 |
| DOI: | 10.1016/j.bbapap.2014.03.008 |
| PMID: | 24657307 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0006-3002 |
|---|---|
| DOI: | 10.1016/j.bbapap.2014.03.008 |