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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Proteoforms of the platelet-aggregating enzyme PA-BJ, a serine proteinase from Bothrops jararaca venom

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Yamashiro, Edson T. [1] ; Oliveira, Ana K. [1] ; Kitano, Eduardo S. [1] ; Menezes, Milene C. [1] ; Junqueira-de-Azevedo, Inacio L. [1] ; Leme, Adriana F. Paes [2] ; Serrano, Solange M. T. [1]
Total Authors: 7
[1] Inst Butantan, Ctr Toxins Immune Response & Cell Signaling CeTIC, BR-05503000 Sao Paulo - Brazil
[2] Lab Nacl Biociencias LNBio, Campinas, SP - Brazil
Total Affiliations: 2
Document type: Journal article
Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1844, n. 12, p. 2068-2076, DEC 2014.
Web of Science Citations: 5

Snake venoms contain serine proteinases that are functionally similar to thrombin and specifically cleave fibrinogen to convert it into fibrin or activate platelets to aggregation. PA-BJ is a serine proteinase from Bothrops jararaca venom that promotes platelet aggregation and this effect is mediated by the G-coupled protein receptors PAR1 and PAR4. In this study we describe an improved procedure to obtain PA-BJ from B. jararaca venom that uses less chromatographic steps, and, interestingly, results in the isolation of eight proteoforms showing slightly different pls and molecular masses due to variations in their glycosylation levels. The identity of the isolated PA-BJ forms (1-8) was confirmed by mass spectrometry, and they showed similar platelet-activating activity on washed platelet suspensions. N- and O-deglycosylation of PA-BJ 1-8 under denaturing conditions generated variable electrophoretic profiles and showed that some forms were resistant to complete deglycosylation. Furthermore, N- and O-deglycosylation under non-denaturing conditions also showed different electrophoretic profiles between the PA-BJ forms and caused partial loss of their ability to cleave a recombinant exodomain of PAR1 receptor. In parallel, three cDNAs encoding PA-BJ-like enzymes were identified by pyrosequencing of a B. jararaca venom gland library constructed with RNA from a single specimen. Taken together, our results suggest that PA-BJ occurs in the B. jararaca venom in multiple proteoforms displaying similar properties upon platelets regardless of their variable isoelectric points, molecular masses, carbohydrate moieties and susceptibility to the activity of glycosidases, and highlight that variability of specific venom components contributes to venom proteome complexity. (C) 2014 Elsevier B.V. All rights reserved. (AU)

FAPESP's process: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling
Grantee:Hugo Aguirre Armelin
Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
FAPESP's process: 11/11308-0 - Proteomic analysis of leaves of sugarcane cultivars and genotypes
Grantee:Eduardo Shigueo Kitano
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 11/08514-8 - Study of the degradome of HF3, a PIII-class snake venom metalloproteinase from Bothrops jararaca venom, on cultured fibroblasts
Grantee:André Zelanis Palitot Pereira
Support Opportunities: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 98/14307-9 - Center for Applied Toxinology
Grantee:Hugo Aguirre Armelin
Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC