Selective oxidation of Met-192 in bovine α-chymotrypsin. Effect on catalytic and inhibitor binding properties

Cutruzzolà, Francesca; Ascenzi, Paolo; Barra, Donatella; Bolognesi, Martino; Menegatti, Enea; Sarti, Paolo; Schnebli, Hans Peter; Tomova, Svetlana; Amiconi, Gino

doi:10.1016/0167-4838(93)90214-c

Cited by 7 publications

(29 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Over the whole pH range explored (i.e., from pH 4.0 to 9.0), the effect of pH on the affinity of native BPTI and [Hse lactone-52]-52,53-seco-BPTI for kallikrein as well as on the values of K, for native BPTI association to chymotrypsin (Fig. 1) is reminiscent of that observed for the binding of (macro)molecular inhibitors to serine (pro)enzymes (Antonini et al, 1983;Menegatti et al, 1984;Amiconi et al, 1987;Ascenzi et al, 1988Ascenzi et al, , 1992Coletta et al, 1990;Onesti et al, 1992;Cutruzzola et al, 1993). On the other hand, a more complex pH effect modulating [Hse lactone-52]-52,53-seco-BPTI binding to chymotrypsin is observed ( Fig.…”

Section: Resultsmentioning

confidence: 74%

“…1 (B)) (Antonini et al, 1983;Menegatti et al, 1984;Amiconi et al, 1987;Ascenzi et al, 1988Coletta et al, 1990;Onesti et al, 1992;Cutruzzola et al, 1993). According to linkage relations (Wyman, 1964;Antonini et al, 1983;Menegatti el al., 1984;Amiconi el al., 1987;Ascenzi et al, 1988Ascenzi et al, , 1992Coletta et al, 1990;Onesti et al, 1992;Cutruzzola et al, 1993), the models holding for native BPTI and [Hse lactone-52]-52,53-seco-BPTI binding to kallikrein and chymotrypsin lead to the following expression:…”

Section: Resultsmentioning

confidence: 99%

“…According to the literature (for reviews see Antonini et al, 1983;Menegatti et al, 1984;Ascenzi et al, 1992;Amiconi et al, 1987;Onesti et al, 1992;Cutruzzola et al, 1993), the increase of K, values for native BPTI and [Hse lactone-52]-52,53-seco-BPTI binding to kallikrein as well as for chymotrypsin : native BPTI complex formation, upon pH increasing from 4.0 to 9.0, may be interpreted as reflecting the acidic pK shift of the proteinase His57 catalytic residue, upon inhibitor binding, from pKLNL=7.1, in the free enzyme, to pKL,, 1 : 5.1, in the proteinase : inhibitor complexes (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

“…Values of the association equilibrium constant ( K a ) for inhibitor binding to kallikrein and chymotrypsin were evaluated by the inhibitory effect of the native BPTI and [Hse lactone-52]-52,53-seco-BPTI on the hydrolysis of ZLysONp catalysed by kallikrein as well as on the cleavage of ZLysONp and ZTyrONp catalysed by chymotrypsin. The proteinase catalyzed hydrolysis of ZLysONp and ZTyrONp was followed spectrophotometrically at 360 nm (Ascenzi et al, 1980(Ascenzi et al, , 1988Antonini el al., 1983;Menegatti et al, 1984;Coletta et al, 1990;Onesti et al, 1992;Cutruzzola et al, 1993). Values of K , were obtained using the graphical method proposed by Ascenzi et al (1987).…”

Section: Methodsmentioning

confidence: 99%

“…Values of K , were obtained using the graphical method proposed by Ascenzi et al (1987). A standard deviation of k8% was evaluated for values of K , (Antonini et al, 1983;Menegatti et al, 1984;Ascenzi et al, 1988Ascenzi et al, , 1992Coletta et af., 1990;Onesti et al, 1992;Cutruzzola et al, 1993).…”

Section: Methodsmentioning

confidence: 99%

See 4 more Smart Citations

Binding of native and [homoserine lactone‐52]‐52,53‐seco‐bovine basic pancreatic trypsin inhibitor (kunitz inhibitor) to porcine pancreatic β‐kallikrein‐B and bovine α‐chymtorpsin: Thermodynaic study

Oddone¹,

Barra²,

Amiconi³

et al. 1994

J of Molecular Recognition

Self Cite

View full text Add to dashboard Cite

Values of the association equilibrium constant (Ka) for the binding of the native and of the cyanogen bromide-cleaved bovine basic pancreatic trypsin inhibitor (native BPTI and [Hse lactone-52]-52,53-seco-BPTI, respectively) to neuraminidase-treated porcine pancreatic beta-kallikrein-B (kallikrein) and bovine alpha-chymotrypsin (chymotrypsin) have been determined between pH 4.0 and 9.0, at 20.0 degrees C. Over the whole pH range explored, native BPTI and [Hse lactone-52]-52,53-seco-BPTI show the same affinity for kallikrein. On the other hand, the affinity of [Hse lactone-52]-52,53-seco-BPTI for chymotrypsin is higher, around neutrality, than that found for native BPTI by about one order of magnitude, converging in the acidic pH limb. The simplest mechanism accounting for the observed data implies that, on lowering the pH from 9.0 to 4.0, (i) the decrease in affinity for the binding of native BPTI to kallikrein and chymotrypsin, as well as for the association of [Hse lactone-52]-52,53-seco-BPTI to kallikrein, reflects the acidic pK shift, upon inhibitor association, of a single ionizing group; and (ii) the decrease of Ka values for [Hse lactone-52]-52,53-seco-BPTI binding to chymotrypsin appears to be modulated by the acidic pK shift, upon inhibitor association, of two non-equivalent proton-binding residues. On the basis of the stereochemistry of the serine proteinase/inhibitor contact region(s), these data indicate that long-range structural changes in [Hse lactone-52]-52,53-seco-BPTI are energetically linked to the chymotrypsin:inhibitor complex formation. This observation represents an important aspect for the mechanism of molecular recognition and regulation in BPTI.

show abstract

Section: Resultsmentioning

confidence: 74%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Binding of native and [homoserine lactone‐52]‐52,53‐seco‐bovine basic pancreatic trypsin inhibitor (kunitz inhibitor) to porcine pancreatic β‐kallikrein‐B and bovine α‐chymtorpsin: Thermodynaic study

Oddone¹,

Barra²,

Amiconi³

et al. 1994

J of Molecular Recognition

Self Cite

View full text Add to dashboard Cite

show abstract

Crystal structure of the bovine α-chymotrypsin:kunitz inhibitor complex. An example of multiple protein:protein recognition sites

et al. 1997

Self Cite

View full text Add to dashboard Cite

The crystal structure of bovine alpha-chymotrypsin (alpha-CHT) in complex with the bovine basic pancreatic trypsin inhibitor (BPTI) has been solved and refined at 2.8 A resolution (R-factor = 0.18). The proteinase:inhibitor complex forms a compact dimer (two alpha-CHT and two BPTI molecules), which may be stabilized by surface-bound sulphate ions, in the crystalline state. Each BPTI molecule, at opposite ends, is contacting both proteinase molecules in the dimer, through the reactive site loop and through residues next to the inhibitor's C-terminal region. Specific recognition between alpha-CHT and BPTI occurs at the (re)active site interface according to structural rules inferred from the analysis of homologous serine proteinase:inhibitor complexes. Lys15, the P1 residue of BPTI, however, does not occupy the alpha-CHT S1 specificity pocket, being hydrogen bonded to backbone atoms of the enzyme surface residues Gly216 and Ser217.

show abstract

Selective oxidation of methionyl residues in the human recombinant secretory leukocyte proteinase inhibitor. Effect on the inhibitor binding properties

Tomova¹,

Cutruzzolà²,

Barra³

et al. 1994

J of Molecular Recognition

Self Cite

View full text Add to dashboard Cite

Binding of the human recombinant secretory leukocyte proteinase inhibitor (SLPI) [native and with the methionyl residues at positions 73, 82, 94 and 96 of domain 2 oxidized to the sulfoxide derivative (Met(O) SLPI)] to bovine alpha-chymotrypsin (alpha-chymotrypsin) [native and with the Met192 residue converted to the sulfoxide derivative (Met(O) alpha-chymotrypsin)] as well as to native bovine beta-trypsin (beta-trypsin), which does not contain methionyl residues, has been investigated between pH 4.0 and 8.0, and between 10.0 degrees C and 30.0 degrees C, from thermodynamic and/or kinetic viewpoints. By increasing the number of oxidized methionyl residues present at the proteinase:inhibitor contact interface (from 0 to 3), the adducts investigated are increasingly destabilized and the relaxation time of the complexes into conformers less stable is enhanced. On the other hand, the selective oxidation of methionyl residues of SLPI and alpha-chymotrypsin, by reaction with chloramine T, does not affect the proteinase inhibition recognition mechanism. Therefore, even though conformational changes may occur in the conversion of native SLPI and native alpha-chymotrypsin to their Met(O) derivatives, a localized steric hindrance can be considered as the main structural determinant accounting for the reported results.

show abstract

Selective oxidation of Met-192 in bovine α-chymotrypsin. Effect on catalytic and inhibitor binding properties

Cited by 7 publications

References 37 publications

Binding of native and [homoserine lactone‐52]‐52,53‐seco‐bovine basic pancreatic trypsin inhibitor (kunitz inhibitor) to porcine pancreatic β‐kallikrein‐B and bovine α‐chymtorpsin: Thermodynaic study

Binding of native and [homoserine lactone‐52]‐52,53‐seco‐bovine basic pancreatic trypsin inhibitor (kunitz inhibitor) to porcine pancreatic β‐kallikrein‐B and bovine α‐chymtorpsin: Thermodynaic study

Crystal structure of the bovine α-chymotrypsin:kunitz inhibitor complex. An example of multiple protein:protein recognition sites

Selective oxidation of methionyl residues in the human recombinant secretory leukocyte proteinase inhibitor. Effect on the inhibitor binding properties

Contact Info

Product

Resources

About