Stopped-flow kinetics of the resynthesis of the reactive site peptide bond in kallikrein inhibitor (Kunitz) by β-trypsin

Quast, Ulrich; Engel, Juergen; Steffen, Erna; Tschesche, Harald; Kupfer, Sigrid

doi:10.1021/bi00602a015

Cited by 54 publications

(30 citation statements)

References 26 publications

(30 reference statements)

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“…It was characterized as described before [12,14]. The molar absorption coefficient was E~~~ = 5.4 x lo3 M -cm-'.…”

Section: Methodsmentioning

confidence: 99%

“…The molar absorption coefficient was E~~~ = 5.4 x lo3 M -cm-'. The inhibitor was converted to I* according to Jering and Tschesche [15,16] and I*-OMe was prepared by esterification of I* with methanol [14]. Both materials were purified and characterized as described earlier [14].…”

Section: Methodsmentioning

confidence: 99%

“…The inhibitor was converted to I* according to Jering and Tschesche [15,16] and I*-OMe was prepared by esterification of I* with methanol [14]. Both materials were purified and characterized as described earlier [14]. Absorption coefficients = 6.1 x lo3 M-' cm-I for I* and 7 x lo3 M-' cm-' for I*-OMe [14].…”

Section: Methodsmentioning

confidence: 99%

“…Both materials were purified and characterized as described earlier [14]. Absorption coefficients = 6.1 x lo3 M-' cm-I for I* and 7 x lo3 M-' cm-' for I*-OMe [14]. a-Chymotrypsin (3 x crystallized, lot CDS 54C404 from Worthington) was checked for its purity by dodecylsulfate gel electrophoresis.…”

Section: Methodsmentioning

confidence: 99%

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Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Quast¹,

Engel²,

Steffen³

et al. 1978

European Journal of Biochemistry

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Modified trypsin kallikrein inhibitor (I*), with the reactive‐site peptide bond Lys‐15–Ala‐16 split, reacts with α‐chymotrypsin (E) via an intermediate X to the stable tetrahedral complex C: E + I*⇌ X → C. Formation of X constitutes a fast pre‐equilibrium (equilibrium constant Kx= 7 × 10−5 M. association rate constant kx= 4 × 103 M−1 s−1) to the slow reaction X → C (rate constant kc= 2 × 10−3 s−1), all values at pH 7.5. No intermediate X is observed when α‐chymotrypsin reacts with I*‐OMe in which the carboxyl group of Lys‐15 is esterified by methanol. This observation as well as the different pH dependence of the overall association rate constants in the case of I* and I*‐OMe indicate that formation of X precedes formation of the acyl enzyme in the catalytic pathway. The data are compared to the similar results obtained with β‐trypsin and I* or I*‐OMe.

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“…It was characterized as described before [12,14]. The molar absorption coefficient was E~~~ = 5.4 x lo3 M -cm-'.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Quast¹,

Engel²,

Steffen³

et al. 1978

European Journal of Biochemistry

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“…Various structural and functional evidence has been pre~ented that the residues preceding Gly 9 of chicken cystatin or ~he homologous Gly ~ in cystatin C bind in the putative $2 and ~' ;3 subsites of papain and are essential for effective inhibition I 18 -20,22]. These findings suggest a formal analogy of the Gly 9-\la 1° bond of chicken cystatin with the 'scissile bond' in the eactive site of small serine proteinase inhibitors, which is fre-, luently cleaved in the enzyme-inhibitor complex according to he so called 'standard mechanism' [5,23,24]. Structural data do lot support this analogy, however: in the docking model of the )apain-chicken cystatin complex and in the experimental strucure of the stefin B-papain complex, the Gly9-Ala ~° bond is ,patially removed from the catalytic Cys 25 residue of papain md thus seems to be not cleavable [25][26][27].…”

Section: Inhibitor Variants As Substratesmentioning

confidence: 99%

Temporary inhibition of papain by hairpin loop mutants of chicken cystatin Distorted binding of the loops results in cleavage of the Gly⁹‐Ala¹⁰ bond

Machleidt¹,

Nägler

Assfalg-Machleidt³

et al. 1995

FEBS Letters

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„Chemische Mutation”︁ durch Aminosäureaustausch im reaktiven Zentrum eines Proteinase‐Inhibitors und Ånderung seiner Hemmspezifität

Wenzel

Tschesche

1981

Angewandte Chemie

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Durch chemische Eingriffe in das aktive Zentrum eines Enzyms ist es gelungen, Lysin gegen Valin, d. h. gegen eine nicht basische Aminosäure, auszutauschen. Auf enzymatischem Wege ist das nicht möglich. Der native Kunitz‐Inhibitor (1) aus Rinderorganen hemmt Trypsin sehr stark und leukocytäre Elastase kaum; das chemisch modifizierte Produkt (2) ist ein sehr guter Inhibitor für Elastase aus menschlichen Leukocyten. Die Methode ist verallgemeinerungsfähigmagnified image.

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Stopped-flow kinetics of the resynthesis of the reactive site peptide bond in kallikrein inhibitor (Kunitz) by β-trypsin

Cited by 54 publications

References 26 publications

Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Temporary inhibition of papain by hairpin loop mutants of chicken cystatin Distorted binding of the loops results in cleavage of the Gly⁹‐Ala¹⁰ bond

„Chemische Mutation”︁ durch Aminosäureaustausch im reaktiven Zentrum eines Proteinase‐Inhibitors und Ånderung seiner Hemmspezifität

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Stopped-flow kinetics of the resynthesis of the reactive site peptide bond in kallikrein inhibitor (Kunitz) by β-trypsin

Cited by 54 publications

References 26 publications

Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Kinetics of the Interaction of α‐Chymotrypsin with Trypsin Kallikrein Inhibitor (Kunitz) in Which the Reactive‐Site Peptide Bond Lys‐15–Ala‐16 is Split

Temporary inhibition of papain by hairpin loop mutants of chicken cystatin Distorted binding of the loops results in cleavage of the Gly9‐Ala10 bond

„Chemische Mutation”︁ durch Aminosäureaustausch im reaktiven Zentrum eines Proteinase‐Inhibitors und Ånderung seiner Hemmspezifität

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Temporary inhibition of papain by hairpin loop mutants of chicken cystatin Distorted binding of the loops results in cleavage of the Gly⁹‐Ala¹⁰ bond