Inhibition of serine proteinases by tetra-p-amidinophenoxy-neo-pentane: Thermodynamic and molecular modeling study

Enea Menegatti, Roberto Ferroni, Santo Scalia, Mario Guarneri, Martino Bolognesi, Paolo Ascenzi, Gino Amiconi

Research output: Contribution to journalArticle

Abstract

The inhibitory effect of the aromatic tetra-benzamidine derivative tetra-p-amidinophenoxy-neo-pentane (TAPP) on the catalytic properties of βtrypsin (EC 3.4.21.4), αthrombin (EC 3.4.21.5), factor Xa (EC 3.4.21.6), Lys77-plasmin (EC 3.4.21.7) and βkallikrein-B (EC 3.4.21.35) was investigated (between pH 2 and 8, I = 0.1 M; T = 37 ± 0.5°C), and analyzed in parallel with that of benzamidine, commonly taken as a molecular inhibitor model of serine proteinases. Over the whole pH range explored, TAPP and benzamidine show the same values of the dissociation inhibition constant (Ki) for βtrypsin; at variance with the affinity of TAPP for αthrombin, factor Xa, Lys77-plasmin and β-kallikrein-B which is higher than that found for benzamidine association around neutrality, but tends to converge in the acidic pH limb. On lowering the pH from 5.5 to 3.0, values of Ki for TAPP binding to β-trypsin as well as for benzamidine association to all the enzymes investigated decreased thus reflecting the pK-shift, upon inhibitor binding, of a single ionizing group. Over the same pH range, values of Ki for TAPP binding to αthrombin, factor Xa, Lys77- plasmin and β-kallikrein-B may be described as depending on the pK-shift, upon inhibitor association, of two equivalent proton-binding amino acid residues. Considering the X-ray three-dimensional structures and the computer-generated molecular models of serine proteinases:TAPP and :benza-midine adducts, the observed binding behaviour of TAPP and benzamidine to the enzymes considered has been related to the inferred stereochemistry of proteinase: inhibitor contact region(s).

Original languageEnglish
Pages (from-to)23-30
Number of pages8
JournalJournal of Enzyme Inhibition and Medicinal Chemistry
Volume2
Issue number1
DOIs
Publication statusPublished - 1987

Fingerprint

Molecular modeling
Serine Proteases
Thermodynamics
Factor Xa
Kallikreins
Fibrinolysin
Thrombin
Trypsin
Molecular Models
Association reactions
Stereochemistry
Serine Proteinase Inhibitors
Enzymes
1,3-di-(4-amidinophenoxy)-2,2-bis-(4-amidinophenoxymethyl)propane
benzamidine
Protons
Peptide Hydrolases
Extremities
X-Rays
Derivatives

Keywords

  • Benzamidine
  • Inhibitor adduct formation (thermodynamics of)
  • Inhibitor adducts (molecular modeling of)
  • Serine proteinase
  • Serine proteinase inhibitors
  • Tetra-p-amidinophenoxy-neo-pentane

ASJC Scopus subject areas

  • Drug Discovery
  • Pharmacology
  • Biochemistry
  • Molecular Medicine

Cite this

Inhibition of serine proteinases by tetra-p-amidinophenoxy-neo-pentane : Thermodynamic and molecular modeling study. / Menegatti, Enea; Ferroni, Roberto; Scalia, Santo; Guarneri, Mario; Bolognesi, Martino; Ascenzi, Paolo; Amiconi, Gino.

In: Journal of Enzyme Inhibition and Medicinal Chemistry, Vol. 2, No. 1, 1987, p. 23-30.

Research output: Contribution to journalArticle

Menegatti, Enea ; Ferroni, Roberto ; Scalia, Santo ; Guarneri, Mario ; Bolognesi, Martino ; Ascenzi, Paolo ; Amiconi, Gino. / Inhibition of serine proteinases by tetra-p-amidinophenoxy-neo-pentane : Thermodynamic and molecular modeling study. In: Journal of Enzyme Inhibition and Medicinal Chemistry. 1987 ; Vol. 2, No. 1. pp. 23-30.
@article{13d3bf7dd3db44a3a027bb761890d14d,
title = "Inhibition of serine proteinases by tetra-p-amidinophenoxy-neo-pentane: Thermodynamic and molecular modeling study",
abstract = "The inhibitory effect of the aromatic tetra-benzamidine derivative tetra-p-amidinophenoxy-neo-pentane (TAPP) on the catalytic properties of βtrypsin (EC 3.4.21.4), αthrombin (EC 3.4.21.5), factor Xa (EC 3.4.21.6), Lys77-plasmin (EC 3.4.21.7) and βkallikrein-B (EC 3.4.21.35) was investigated (between pH 2 and 8, I = 0.1 M; T = 37 ± 0.5°C), and analyzed in parallel with that of benzamidine, commonly taken as a molecular inhibitor model of serine proteinases. Over the whole pH range explored, TAPP and benzamidine show the same values of the dissociation inhibition constant (Ki) for βtrypsin; at variance with the affinity of TAPP for αthrombin, factor Xa, Lys77-plasmin and β-kallikrein-B which is higher than that found for benzamidine association around neutrality, but tends to converge in the acidic pH limb. On lowering the pH from 5.5 to 3.0, values of Ki for TAPP binding to β-trypsin as well as for benzamidine association to all the enzymes investigated decreased thus reflecting the pK-shift, upon inhibitor binding, of a single ionizing group. Over the same pH range, values of Ki for TAPP binding to αthrombin, factor Xa, Lys77- plasmin and β-kallikrein-B may be described as depending on the pK-shift, upon inhibitor association, of two equivalent proton-binding amino acid residues. Considering the X-ray three-dimensional structures and the computer-generated molecular models of serine proteinases:TAPP and :benza-midine adducts, the observed binding behaviour of TAPP and benzamidine to the enzymes considered has been related to the inferred stereochemistry of proteinase: inhibitor contact region(s).",
keywords = "Benzamidine, Inhibitor adduct formation (thermodynamics of), Inhibitor adducts (molecular modeling of), Serine proteinase, Serine proteinase inhibitors, Tetra-p-amidinophenoxy-neo-pentane",
author = "Enea Menegatti and Roberto Ferroni and Santo Scalia and Mario Guarneri and Martino Bolognesi and Paolo Ascenzi and Gino Amiconi",
year = "1987",
doi = "10.3109/14756368709030353",
language = "English",
volume = "2",
pages = "23--30",
journal = "Journal of Enzyme Inhibition and Medicinal Chemistry",
issn = "1475-6366",
publisher = "Taylor and Francis Ltd.",
number = "1",

}

TY - JOUR

T1 - Inhibition of serine proteinases by tetra-p-amidinophenoxy-neo-pentane

T2 - Thermodynamic and molecular modeling study

AU - Menegatti, Enea

AU - Ferroni, Roberto

AU - Scalia, Santo

AU - Guarneri, Mario

AU - Bolognesi, Martino

AU - Ascenzi, Paolo

AU - Amiconi, Gino

PY - 1987

Y1 - 1987

N2 - The inhibitory effect of the aromatic tetra-benzamidine derivative tetra-p-amidinophenoxy-neo-pentane (TAPP) on the catalytic properties of βtrypsin (EC 3.4.21.4), αthrombin (EC 3.4.21.5), factor Xa (EC 3.4.21.6), Lys77-plasmin (EC 3.4.21.7) and βkallikrein-B (EC 3.4.21.35) was investigated (between pH 2 and 8, I = 0.1 M; T = 37 ± 0.5°C), and analyzed in parallel with that of benzamidine, commonly taken as a molecular inhibitor model of serine proteinases. Over the whole pH range explored, TAPP and benzamidine show the same values of the dissociation inhibition constant (Ki) for βtrypsin; at variance with the affinity of TAPP for αthrombin, factor Xa, Lys77-plasmin and β-kallikrein-B which is higher than that found for benzamidine association around neutrality, but tends to converge in the acidic pH limb. On lowering the pH from 5.5 to 3.0, values of Ki for TAPP binding to β-trypsin as well as for benzamidine association to all the enzymes investigated decreased thus reflecting the pK-shift, upon inhibitor binding, of a single ionizing group. Over the same pH range, values of Ki for TAPP binding to αthrombin, factor Xa, Lys77- plasmin and β-kallikrein-B may be described as depending on the pK-shift, upon inhibitor association, of two equivalent proton-binding amino acid residues. Considering the X-ray three-dimensional structures and the computer-generated molecular models of serine proteinases:TAPP and :benza-midine adducts, the observed binding behaviour of TAPP and benzamidine to the enzymes considered has been related to the inferred stereochemistry of proteinase: inhibitor contact region(s).

AB - The inhibitory effect of the aromatic tetra-benzamidine derivative tetra-p-amidinophenoxy-neo-pentane (TAPP) on the catalytic properties of βtrypsin (EC 3.4.21.4), αthrombin (EC 3.4.21.5), factor Xa (EC 3.4.21.6), Lys77-plasmin (EC 3.4.21.7) and βkallikrein-B (EC 3.4.21.35) was investigated (between pH 2 and 8, I = 0.1 M; T = 37 ± 0.5°C), and analyzed in parallel with that of benzamidine, commonly taken as a molecular inhibitor model of serine proteinases. Over the whole pH range explored, TAPP and benzamidine show the same values of the dissociation inhibition constant (Ki) for βtrypsin; at variance with the affinity of TAPP for αthrombin, factor Xa, Lys77-plasmin and β-kallikrein-B which is higher than that found for benzamidine association around neutrality, but tends to converge in the acidic pH limb. On lowering the pH from 5.5 to 3.0, values of Ki for TAPP binding to β-trypsin as well as for benzamidine association to all the enzymes investigated decreased thus reflecting the pK-shift, upon inhibitor binding, of a single ionizing group. Over the same pH range, values of Ki for TAPP binding to αthrombin, factor Xa, Lys77- plasmin and β-kallikrein-B may be described as depending on the pK-shift, upon inhibitor association, of two equivalent proton-binding amino acid residues. Considering the X-ray three-dimensional structures and the computer-generated molecular models of serine proteinases:TAPP and :benza-midine adducts, the observed binding behaviour of TAPP and benzamidine to the enzymes considered has been related to the inferred stereochemistry of proteinase: inhibitor contact region(s).

KW - Benzamidine

KW - Inhibitor adduct formation (thermodynamics of)

KW - Inhibitor adducts (molecular modeling of)

KW - Serine proteinase

KW - Serine proteinase inhibitors

KW - Tetra-p-amidinophenoxy-neo-pentane

UR - http://www.scopus.com/inward/record.url?scp=0023639781&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023639781&partnerID=8YFLogxK

U2 - 10.3109/14756368709030353

DO - 10.3109/14756368709030353

M3 - Article

C2 - 3508169

AN - SCOPUS:0023639781

VL - 2

SP - 23

EP - 30

JO - Journal of Enzyme Inhibition and Medicinal Chemistry

JF - Journal of Enzyme Inhibition and Medicinal Chemistry

SN - 1475-6366

IS - 1

ER -