Proteinase inhibitors from desert locust, Schistocerca gregaria: Engineering of both P1 and P1' residues converts a potent chymotrypsin inhibitor to a potent trypsin inhibitor

Zulfiquar Malik, Sumaira Amir, Gábor Pál, Zsuzsa Buzás, Éva Várallyay, József Antal, Zoltán Szilágyi, Károly Vékey, Bence Asbóth, András Patthy, László Gráf

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Two peptides, SGCI and SGTI, that inhibited chymotrypsin and trypsin, respectively, were isolated from the haemolymph of Schistocerca gregaria. Their primary structures were found to be identical with SGP-2 and SGP-1, two of a series of peptides isolated from ovaries of the same species (A. Hamdaoui et al., FEBS Lett. 422 (1998) 74-78). All these peptides are composed of 35-36 amino acid residues and contain three homologous disulfide bridges. The residues imparting specificity to SGCI and SGTI were identified as Leu-30 and Arg-29, respectively. The peptides were synthesised by solid- phase peptide synthesis, and the synthetic ones displayed the same inhibition as the natural forms: SGCI is a strong inhibitor of chymotrypsin (K(i)=6.2 x 10-12 M), and SGTI is a rather weak inhibitor of trypsin (K(i)=2.1 x 10- 7 M). The replacement of P1 then P1' residues of SGCI with trypsin- specific residues increased affinity towards trypsin 3600- and 1100-fold, respectively, thus SGCI was converted to a strong trypsin inhibitor (K(i)=5.0 x 10-12 M) that retained some inhibitory affinity towards chymotrypsin (K(i)=3.5 x 10-8 M). The documented role of both P1 and P1' highlights the importance of S1'P1' interactions in enzyme-inhibitor complexes.

Original languageEnglish (US)
Pages (from-to)143-150
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1434
Issue number1
DOIs
StatePublished - Sep 14 1999
Externally publishedYes

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Grasshoppers
Trypsin Inhibitors
Chymotrypsin
Peptide Hydrolases
Peptides
Trypsin
Solid-Phase Synthesis Techniques
Hemolymph
Enzyme Inhibitors
Disulfides
Ovary
Amino Acids
isoinhibitor K

Keywords

  • Chymotrypsin
  • Insect peptide
  • Protein protease inhibitor
  • Schistocerca gregaria
  • Subsite specificity

ASJC Scopus subject areas

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Proteinase inhibitors from desert locust, Schistocerca gregaria : Engineering of both P1 and P1' residues converts a potent chymotrypsin inhibitor to a potent trypsin inhibitor. / Malik, Zulfiquar; Amir, Sumaira; Pál, Gábor; Buzás, Zsuzsa; Várallyay, Éva; Antal, József; Szilágyi, Zoltán; Vékey, Károly; Asbóth, Bence; Patthy, András; Gráf, László.

In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, Vol. 1434, No. 1, 14.09.1999, p. 143-150.

Research output: Contribution to journalArticle

Malik, Zulfiquar ; Amir, Sumaira ; Pál, Gábor ; Buzás, Zsuzsa ; Várallyay, Éva ; Antal, József ; Szilágyi, Zoltán ; Vékey, Károly ; Asbóth, Bence ; Patthy, András ; Gráf, László. / Proteinase inhibitors from desert locust, Schistocerca gregaria : Engineering of both P1 and P1' residues converts a potent chymotrypsin inhibitor to a potent trypsin inhibitor. In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. 1999 ; Vol. 1434, No. 1. pp. 143-150.
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abstract = "Two peptides, SGCI and SGTI, that inhibited chymotrypsin and trypsin, respectively, were isolated from the haemolymph of Schistocerca gregaria. Their primary structures were found to be identical with SGP-2 and SGP-1, two of a series of peptides isolated from ovaries of the same species (A. Hamdaoui et al., FEBS Lett. 422 (1998) 74-78). All these peptides are composed of 35-36 amino acid residues and contain three homologous disulfide bridges. The residues imparting specificity to SGCI and SGTI were identified as Leu-30 and Arg-29, respectively. The peptides were synthesised by solid- phase peptide synthesis, and the synthetic ones displayed the same inhibition as the natural forms: SGCI is a strong inhibitor of chymotrypsin (K(i)=6.2 x 10-12 M), and SGTI is a rather weak inhibitor of trypsin (K(i)=2.1 x 10- 7 M). The replacement of P1 then P1' residues of SGCI with trypsin- specific residues increased affinity towards trypsin 3600- and 1100-fold, respectively, thus SGCI was converted to a strong trypsin inhibitor (K(i)=5.0 x 10-12 M) that retained some inhibitory affinity towards chymotrypsin (K(i)=3.5 x 10-8 M). The documented role of both P1 and P1' highlights the importance of S1'P1' interactions in enzyme-inhibitor complexes.",
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AU - Amir, Sumaira

AU - Pál, Gábor

AU - Buzás, Zsuzsa

AU - Várallyay, Éva

AU - Antal, József

AU - Szilágyi, Zoltán

AU - Vékey, Károly

AU - Asbóth, Bence

AU - Patthy, András

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AB - Two peptides, SGCI and SGTI, that inhibited chymotrypsin and trypsin, respectively, were isolated from the haemolymph of Schistocerca gregaria. Their primary structures were found to be identical with SGP-2 and SGP-1, two of a series of peptides isolated from ovaries of the same species (A. Hamdaoui et al., FEBS Lett. 422 (1998) 74-78). All these peptides are composed of 35-36 amino acid residues and contain three homologous disulfide bridges. The residues imparting specificity to SGCI and SGTI were identified as Leu-30 and Arg-29, respectively. The peptides were synthesised by solid- phase peptide synthesis, and the synthetic ones displayed the same inhibition as the natural forms: SGCI is a strong inhibitor of chymotrypsin (K(i)=6.2 x 10-12 M), and SGTI is a rather weak inhibitor of trypsin (K(i)=2.1 x 10- 7 M). The replacement of P1 then P1' residues of SGCI with trypsin- specific residues increased affinity towards trypsin 3600- and 1100-fold, respectively, thus SGCI was converted to a strong trypsin inhibitor (K(i)=5.0 x 10-12 M) that retained some inhibitory affinity towards chymotrypsin (K(i)=3.5 x 10-8 M). The documented role of both P1 and P1' highlights the importance of S1'P1' interactions in enzyme-inhibitor complexes.

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