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| Format: | Artículo científico |
| Language: | en |
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International journal of biological macromolecules
2026
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41921802/ |
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| author | Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta |
| author_facet | Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta |
| collection | PubMed - marine biology |
| contents | Structural characterization of the serine protease activity of Natterin-like 2 from Thalassophryne maculosa venom. Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta Animals Serine Proteases Amino Acid Sequence Kinetics This study primarily characterizes one of the four Natterin-like subunits (Natterin-like 2) identified in the venom of Thalassophryne maculosa, for which partial sequence information was obtained by mass spectrometry. Peptide fingerprinting revealed sequences similar to those of the four Natterins previously described in T. nattereri, here designated as Natterin-like subunits. The identification of fragments corresponding to four monomeric subunits within a trimer sized band suggests the existence of trimeric biological units with different stoichiometries. All unglycosylated monomeric subunits exhibited alterations in the positioning of the serine protease catalytic triad residues; however, after purification, at least one subunit retained proteolytic activity. Sequence assembly was most complete for Natterin-like 2, which served as the basis for the structural analyses described herein. Inhibition assays guided the use of affinity chromatography to enrich the Natterin-like fraction and determine kinetic parameters. The purified fraction displayed an apparent Km of 39.4 μM with azocasein, approximately 2.5 fold higher than the 15.5 μM measured with BAEE. Both kcat and catalytic efficiency were higher for BAEE than for azocasein, with kcat values of 15.01 s and 10.8 s, and catalytic efficiencies of 9.23x10 M s and 2.7x10 M s, respectively, consistent with trypsin-like activity. Comparative analyses support the conservation of residues nH122, nD146, and nS242 in Natterin-like 2, suggesting a putative catalytic triad analogous to that of serine proteases. Although the trimeric arrangement resembles HtrA proteases, conserved cysteines, benzamidine inhibition, and kinetic parameters indicate a trypsin-like catalytic mechanism for the Natterin-like proteins in T. maculosa venom. |
| format | Artículo científico |
| id | pubmed_41921802 |
| institution | PubMed |
| language | en |
| publishDate | 2026 |
| publisher | International journal of biological macromolecules |
| record_format | pubmed |
| spellingShingle | Structural characterization of the serine protease activity of Natterin-like 2 from Thalassophryne maculosa venom. Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta Animals Serine Proteases Amino Acid Sequence Kinetics Structural characterization of the serine protease activity of Natterin-like 2 from Thalassophryne maculosa venom. Vásquez-Suárez, Aleikar De Sousa, Leonardo Bolaños-Curvelo, Juan Macaya-Zapata, Luis Peralta, Christian Cronshaw, Andrew Bunster, Marta Animals Serine Proteases Amino Acid Sequence Kinetics This study primarily characterizes one of the four Natterin-like subunits (Natterin-like 2) identified in the venom of Thalassophryne maculosa, for which partial sequence information was obtained by mass spectrometry. Peptide fingerprinting revealed sequences similar to those of the four Natterins previously described in T. nattereri, here designated as Natterin-like subunits. The identification of fragments corresponding to four monomeric subunits within a trimer sized band suggests the existence of trimeric biological units with different stoichiometries. All unglycosylated monomeric subunits exhibited alterations in the positioning of the serine protease catalytic triad residues; however, after purification, at least one subunit retained proteolytic activity. Sequence assembly was most complete for Natterin-like 2, which served as the basis for the structural analyses described herein. Inhibition assays guided the use of affinity chromatography to enrich the Natterin-like fraction and determine kinetic parameters. The purified fraction displayed an apparent Km of 39.4 μM with azocasein, approximately 2.5 fold higher than the 15.5 μM measured with BAEE. Both kcat and catalytic efficiency were higher for BAEE than for azocasein, with kcat values of 15.01 s and 10.8 s, and catalytic efficiencies of 9.23x10 M s and 2.7x10 M s, respectively, consistent with trypsin-like activity. Comparative analyses support the conservation of residues nH122, nD146, and nS242 in Natterin-like 2, suggesting a putative catalytic triad analogous to that of serine proteases. Although the trimeric arrangement resembles HtrA proteases, conserved cysteines, benzamidine inhibition, and kinetic parameters indicate a trypsin-like catalytic mechanism for the Natterin-like proteins in T. maculosa venom. |
| title | Structural characterization of the serine protease activity of Natterin-like 2 from Thalassophryne maculosa venom. |
| topic | Animals Serine Proteases Amino Acid Sequence Kinetics |
| url | https://pubmed.ncbi.nlm.nih.gov/41921802/ |