Chemical Synthesis and Kinetic Study of the Smallest Naturally Occurring Trypsin Inhibitor SFTI-1 Isolated from Sunflower Seeds and Its Analogues

Zabłotna, Ewa; Kaźmierczak, Katarzyna; Jaśkiewicz, Anna; Stawikowski, Maciej; Kupryszewski, G.; Rolka, Krzysztof

doi:10.1006/bbrc.2002.6746

Cited by 69 publications

(108 citation statements)

References 23 publications

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“…It is produced in nature in a cyclic form inhibiting trypsin with subnanomolar equilibrium constant. Previous studies showed that an opened version of SFTI retains near wild-type affinity as long as the disulfide is intact (27,41,42). On the basis of its structure, SFTI mimics the protease interacting loop of the much larger canonical inhibitor scaffold of Bowman-Birk inhibitors (28).…”

Section: Design and Construction Of The Inhibitory Peptide-phage Librarymentioning

confidence: 99%

Selective Inhibition of the Lectin Pathway of Complement with Phage Display Selected Peptides against Mannose-Binding Lectin-Associated Serine Protease (MASP)-1 and -2: Significant Contribution of MASP-1 to Lectin Pathway Activation

Kocsis

Kékesi

Szász

et al. 2010

The Journal of Immunology

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The complement system, an essential part of the innate immune system, can be activated through three distinct routes: the classical, the alternative, and the lectin pathways. The contribution of individual activation pathways to different biological processes can be assessed by using pathway-selective inhibitors. In this paper, we report lectin pathway-specific short peptide inhibitors developed by phage display against mannose-binding lectin-associated serine proteases (MASPs), MASP-1 and MASP-2. On the basis of the selected peptide sequences, two 14-mer peptides, designated as sunflower MASP inhibitor (SFMI)-1 and SFMI-2, were produced and characterized. SFMI-1 inhibits both MASP-1 and MASP-2 with a KI of 65 and 1030 nM, respectively, whereas SFMI-2 inhibits only MASP-2 with a KI of 180 nM. Both peptides block the lectin pathway activation completely while leaving the classical and the alternative routes intact and fully functional, demonstrating that of all complement proteases only MASP-1 and/or MASP-2 are inhibited by these peptides. In a C4 deposition inhibitor assay using preactivated MASP-2, SFMI-2 is 10-fold more effective than SFMI-1 in accordance with the fact that SFMI-2 is a more potent inhibitor of MASP-2. Surprisingly, however, out of the two peptides, SFMI-1 is much more effective in preventing C3 and C4 deposition when normal human serum containing zymogen MASPs is used. This suggests that MASP-1 has a crucial role in the initiation steps of lectin pathway activation most probably by activating MASP-2. Because the lectin pathway has been implicated in several life-threatening pathological states, these inhibitors should be considered as lead compounds toward developing lectin pathway blocking therapeutics.

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Section: Design and Construction Of The Inhibitory Peptide-phage Librarymentioning

confidence: 99%

Selective Inhibition of the Lectin Pathway of Complement with Phage Display Selected Peptides against Mannose-Binding Lectin-Associated Serine Protease (MASP)-1 and -2: Significant Contribution of MASP-1 to Lectin Pathway Activation

Kocsis

Kékesi

Szász

et al. 2010

The Journal of Immunology

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show abstract

“…Cajanus indicus) [125][126][127][128][129][130][131][132][133][134][135][136][137][138][139][140]. It has been found as an effective chemopreventive agent against prostate cancer in animal models by blocking the generation of reactive oxygen species in prostate cancer cells [141].…”

Section: Bowmam-birk Protease Inhibitors (Bbi)mentioning

confidence: 99%

An Insight of Pulses: From Food to Cancer Treatment

Vohra¹,

Dureja²,

Garg³

2016

J Pharmacogn Nat Prod

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“…SFTI-1 is a backbone-cyclized peptide composed of 14 amino acids and bisected by one disulphide bridge into binding and secondary loops. SFTI-1 belongs to the well-characterized Bowman-Birk family of natural inhibitors and displays high affinity towards trypsin (based on the independent colorimetric analyses, the association constant K a = 1.1 9 10 10 M -1 [6] and inhibition constant K i = 0.1 nM [5], 1 nM [7] and 13 nM [8]). Moreover, we showed that both native bicyclic SFTI-1 and its analogue deprived of the cyclic backbone, but having the disulphide bond, have comparable inhibitory activity [6].…”

Section: Introductionmentioning

confidence: 99%

“…SFTI-1 belongs to the well-characterized Bowman-Birk family of natural inhibitors and displays high affinity towards trypsin (based on the independent colorimetric analyses, the association constant K a = 1.1 9 10 10 M -1 [6] and inhibition constant K i = 0.1 nM [5], 1 nM [7] and 13 nM [8]). Moreover, we showed that both native bicyclic SFTI-1 and its analogue deprived of the cyclic backbone, but having the disulphide bond, have comparable inhibitory activity [6]. Bowman-Birk inhibitors interact with their cognate enzymes via the standard mechanism using a common structural motif: a proteasebinding loop [9,10].…”

Section: Introductionmentioning

confidence: 99%

Interactions between trypsin and its peptidic inhibitors studied by isothermal titration calorimetry (ITC)

Dębowski

Wyrzykowski

Lubos

et al. 2015

J Therm Anal Calorim

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Isothermal titration calorimetry (ITC) technique was used to study the interactions of trypsin with bicyclic sunflower-derived trypsin inhibitor (SFTI-1) as well as with its new monocyclic (with disulphide bridge only) analogues (C 3 H 5 O)-SFTI-1 and (C 8 H 15 O)-SFTI-1. ITC measurements were run in 50 mM buffer solution of HEPES or Tricine of pH 8, containing 20 mM CaCl 2 at 298.15 K. Based on calorimetric data, the equilibrium constants for the inhibitor-enzyme-binding processes, K, the binding stoichiometry, N (inhibitor-to-enzyme molar ratio), as well as thermodynamic parameters (DG, DH, DS) for the reactions were determined. The study revealed that the stoichiometry of the resulting complexes equals 1:1. The negative binding enthalpy (D ITC H) and favourable entropy factor (TD ITC S) suggest an important contribution of hydrogen bonding as well as hydrophobic interactions to the inhibitor-enzyme affinity. Furthermore, the relationship between the modification of the peptide structure, the experimental conditions and the thermodynamic parameters has been discussed.

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Chemical Synthesis and Kinetic Study of the Smallest Naturally Occurring Trypsin Inhibitor SFTI-1 Isolated from Sunflower Seeds and Its Analogues

Cited by 69 publications

References 23 publications

Selective Inhibition of the Lectin Pathway of Complement with Phage Display Selected Peptides against Mannose-Binding Lectin-Associated Serine Protease (MASP)-1 and -2: Significant Contribution of MASP-1 to Lectin Pathway Activation

Selective Inhibition of the Lectin Pathway of Complement with Phage Display Selected Peptides against Mannose-Binding Lectin-Associated Serine Protease (MASP)-1 and -2: Significant Contribution of MASP-1 to Lectin Pathway Activation

An Insight of Pulses: From Food to Cancer Treatment

Interactions between trypsin and its peptidic inhibitors studied by isothermal titration calorimetry (ITC)

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