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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2015.tde-04122015-135837
Document
Author
Full name
Roberto Tadashi Kodama
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Portaro, Fernanda Calheta Vieira (President)
Albuquerque, Cristina Northfleet de
Tashima, Alexandre Keiji
Title in Portuguese
Análise dos constituintes de baixa massa molecular de quatro venenos do gênero Bitis e suas atividades biológicas.
Keywords in Portuguese
Bitis
Enzima conversora de angiotensina I
Hipotensão
Peptídeo potencializador de bradicinina (BPP)
Peptídeo rico em prolina
Veneno
Abstract in Portuguese
Na África subsaariana, as serpentes do gênero Bitis são de extrema importância, pois suas vítimas apresentam sintomas como dano local, hemorragia e uma severa hipotensão. Este trabalho identificou moléculas capazes de inibir a atividade da enzima conversora de angiotensina I (ECA) presentes no veneno de quatro serpentes do gênero Bitis. Para isto, as porções de baixa massa molecular desses 4 venenos foram fracionadas em RP-HPLC e as frações com boa inibição sobre a atividade da ECA foram analisadas por espectrometria de massas. Foram identificados 34 oligopeptídeos ricos em prolina (PRO), sendo 8 sintetizados e suas constantes de inibição (Ki) determinadas. Em testes com substratos naturais da ECA, angiotensina I e bradicinina, foi constatada a maior inibição da hidrólise da angiotensina I por quatro PROs. Todos os PROs in vivo reduziram a pressão arterial, e seis deles aumentaram a frequência cardíaca em ratos Wistar. Com isto, conclui-se que existem toxinas no veneno de serpentes do gênero Bitis responsáveis pela hipotensão.
Title in English
Analysis of the low molecular mass constituents from the venom of four species of the Bitis genus and biological activities.
Keywords in English
Bitis
Angiotensin-converting enzyme (ACE)
Bradykinin-potentiating peptide (BPP)
Hypotension
Proline-rich oligpeptide
Venom
Abstract in English
In the sub-saharian Africa, snakes from the Bitis genus are of extreme medical importance, since its victims show symptoms as local tissue damage, hemorrhage and a severe hypotension. This work identified molecules that inhibit the angiotensin I converting enzyme (ACE) in the venom of 4 snakes from the Bitis genus. The low molecular portions of the venom of these snakes were fractionated in RP-HPLC and the fractions that efficiently inhibited the ACE activity were analyzed by mass spectrometry. 34 proline-rich oligopeptides were identified, 8 of them synthesized and had their inhibition constants (Ki) determined. In tests using natural substrates of ACE, angiotensin I and bradykinin, the angiotensin I hydrolysis were better inhibited by four PROs. In vivo tests results showed that all PROs decreased the mean arterial pressure and six of them increased the heart rate. Therefore, we can conclude that there are toxins present in the venom of Bitis capable of cause hypotension.
 
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Publishing Date
2015-12-07
 
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