Hemorrhagic Activity of HF3, a Snake Venom Metalloproteinase: Insights from the Proteomic Analysis of Mouse Skin and Blood Plasma

Abstract

Hemorrhage induced by snake venom metalloproteinases (SVMPs) is a complex phenomenon resulting in capillary disruption and blood extravasation. The mechanism of action of SVMPs has been investigated using various methodologies however the precise molecular events associated with microvessel disruption remains not fully understood. To gain insight into the hemorrhagic process, we analyzed the global effects of HF3, an extremely hemorrhagic SVMP from Bothrops jararaca, in the mouse skin and plasma. We report that in the HF3-treated skin there was evidence of degradation of extracellular matrix (collagens and proteoglycans), cytosolic, cytoskeleton, and plasma proteins. Furthermore, the data suggest that direct and indirect effects promoted by HF3 contributed to tissue injury as the activation of collagenases was detected in the HF3-treated skin. In the plasma analysis after depletion of the 20 most abundant proteins, fibronectin appeared as degraded by HF3. In contrast, some plasma proteinase inhibitors showed higher abundance compared to control skin and plasma. This is the first study to assess the complex in vivo effects of HF3 using high-throughput proteomic approaches, and the results underscore a scenario characterized by the interplay between the hydrolysis of intracellular, extracellular, and plasma proteins and the increase of plasma inhibitors in the hemorrhagic process.

Acidulated Phosphate Fluoride Application Changes the Protein Composition of Human Acquired Enamel Pellicle

“…( table 1 ). Considering the fold change of 1.5 [Aragão et al, 2012a;Paes Leme et al, 2012], amylase has similar abundance among the samples and it was identified in a notable manner with the highest spectral counts, which means a high abundance of this protein in the acquired enamel pellicle independently of the enamel pretreatment. This result is consistent with earlier findings that indicated that amylase is one of the major components of pellicle [Hannig et al, 2004[Hannig et al, , 2005Siqueira et al, 2012b].…”

Proteomic Analysis of Matrix of Dental Biofilm Formed under Dietary Carbohydrate Exposure

“…Differential protein spots were excised, reduced, alkylated and submitted to in-gel digestion with trypsin as described before [Paes Leme et al , 2012]. An aliquot (4.5 l) of the resulting peptide mixture was separated by C18 (75 m !…”

“…The whole extract of proteins (30 g) from each experimental condition was reduced, alkylated and digested with trypsin (Sigma) (1: 50) in 50 m M ammonium bicarbonate for 16 h at 37 ° C. The analysis was done as described before [Paes Leme et al , 2012], except that it was performed over 60 min with two runs for each sample. The resulting fragment spectra were searched as described above with decoy database with a maximum estimated false discovery rate of 1%.…”

Análise proteômica da atividade proteolítica do HF3, uma metaloproteinase do veneno da Bothrops jararaca, no plasma humano e de serpente

“…Ainda que as cadeias alfa e beta fibrinogênio já tenham sido descritas como substratos do HF3 (Oliveira et al, 2010;Paes Leme et al, 2012), neste estudo a cadeia gama do fibrinogênio foi também detectada como um alvo desta metaloproteinase. Peptídeos oriundos da cadeia gama do fibrinogênio foram identificados como produto da atividade do HF3 no P(dep20) e P(enrq) (8)(9).…”

“…No estudo de Paes Leme et al (2012), foi verificado que a apolipoproteína A-II, o segundo maior componente do HDL, foi claramente degradada na pele hemorrágica. No presente estudo foi verificado que não somente a apolipoproteína A-II, como também outras proteínas desta classe (apolipoproteína A-I, A-IV, C-I, C-II, C-III, E, F e L1) foram detectadas como substrato do HF3 (Tabelas 2-9).…”