Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/35939
Title: Substrate specificity studies of the cysteine peptidases falcipain-2 and falcipain-3 from Plasmodium falciparum and demonstration of their kininogenase activity
Authors: Cotrin, Simone Silva [UNIFESP]
Gouvea, Iuri E. [UNIFESP]
Melo, Pollyana M. S. [UNIFESP]
Bagnaresi, Piero [UNIFESP]
Assis, Diego M. [UNIFESP]
Araujo, Mariana S. [UNIFESP]
Juliano, Maria Aparecida [UNIFESP]
Gazarini, Marcos L. [UNIFESP]
Rosenthal, Philip J.
Juliano, Luiz [UNIFESP]
Carmona, Adriana K. [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Univ Calif San Francisco
Keywords: Plasmodium falciparum
Falcipain
FRET substrates
Kinin-releasing
Issue Date: 1-Feb-2013
Publisher: Elsevier B.V.
Citation: Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 187, n. 2, p. 111-116, 2013.
Abstract: We studied the substrate specificity requirements of recombinant cysteine peptidases from Plasmodium falciparum, falcipain-2 (FP-2) and falcipain-3 (FP-3), using fluorescence resonance energy transfer (FRET) peptides as substrates. Systematic modifications were introduced in the lead sequence Abz-KLRSSKQ-EDDnp (Abz = ortho-aminobenzoic acid; EDDnp = N-[2,4-dinitrophenyl]ethylenediamine) resulting in five series assayed to map S-3 - S-2' subsite specificity. Despite high sequence identity and structural similarity between FP-2 and FP-3, noteworthy differences in substrate specificity were observed. the S-1 subsite of FP-2 preferentially accommodates peptides containing the positively charged residue Arg in P-1, while FP-3 has a clear preference for the hydrophobic residue Leu in this position. the S-2 subsite of FP-2 and FP-3 presents a strict specificity for hydrophobic residues, with Leu being the residue preferred by both enzymes. FP-2 did not show preference for the residues present at P-3, while FP-3 hydrolysed the peptide Abz-ALRSSRQ-EDDnp, containing Ala at P-3, with the highest catalytic efficiency of all series studied. FP-2 has high susceptibility for substrates containing hydrophobic residues in P-1', while FP-3 accommodates well peptides containing Arg in this position. the S-2' subsite of both enzymes demonstrated broad specificity. in addition, radioimmunoassay experiments indicated that kinins can be generated by FP-2 and FP-3 proteolysis of high molecular weight kininogen (HK). Both enzymes excised Met-Lys-bradykinin, Lys-bradykinin and bradykinin from the fluorogenic peptide Abz-MISLMKRPPGFSPFRSSRI-NH2, which corresponds to the Met(375) to Ile(393) sequence of HK. the capability of FP-2 and FP-3 to release kinins suggests the involvement of these enzymes in the modulation of malaria pathophysiology. (c) 2013 Elsevier B.V. All rights reserved.
URI: http://repositorio.unifesp.br/handle/11600/35939
ISSN: 0166-6851
Other Identifiers: http://dx.doi.org/10.1016/j.molbiopara.2013.01.002
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