Biocomputing and biochemical studies of enzymes - Manzetti, Sergio
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The human kallikrein family is a family of proteolytic enzymes, classified as serine proteases, that derive from chromosome 19, locus 13.3-13.4. One member of the family, human kallikrein 4 (hK4) which is thought to carry out crucial functions in the prostate, was expressed in this study as a secreted protein in a baculovirus expression system, bearing a His-tag and V5-epitope that were used for purification and detection respectively. Computational analysis was included by applying high-performing computing techniques, such as molecular dynamics, and flexible ligand docking, to predict…mehr

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Produktbeschreibung
The human kallikrein family is a family of proteolytic enzymes, classified as serine proteases, that derive from chromosome 19, locus 13.3-13.4. One member of the family, human kallikrein 4 (hK4) which is thought to carry out crucial functions in the prostate, was expressed in this study as a secreted protein in a baculovirus expression system, bearing a His-tag and V5-epitope that were used for purification and detection respectively. Computational analysis was included by applying high-performing computing techniques, such as molecular dynamics, and flexible ligand docking, to predict antigenic regions, the likely substrate specificity and putative inhibitors. These results show that hK4 has a loop, between Leu83-Ser94 that shows promise as a specific segment that can be exploited for generation of antibodies. The principles of substrate-specificity analysis that were developed were further applied on three metzincins, MMP-3, ADAM-9 and ADAM-10. These three enzymes are metalloproteases, which are involved in tissue remodeling, intracellular signalling and cell-to-cell mediation. This study is a demostration of the power of combining bioinformatics + wet-lab biochemistry.
Autorenporträt
Sergio recevied his Master''s Degree at Queensland Univestiy of Technology - Australia in molecular biology, biotechnology and structural bioinformatics. Sergio Manzetti administers today his own research institute at Fjordforsk.com which publishes studies in several areas, from nanotechnology and environmental sciences to quantum chemistry.