8d49ae6fa63c1189913cd33f5682377a mardru_a2019v17n9p511.pdf 0f56278c6dcb5399afda1c83e3fc67a07f587d80 mardru_a2019v17n9p511.pdf ed80457cdab6ab4fd4f57b1a958acffa3f6c7a514af83caa760d1a3f8c0f1967 mardru_a2019v17n9p511.pdf Title: Characterization, Recombinant Production and Structure-Function Analysis of NvCI, A Picomolar Metallocarboxypeptidase Inhibitor from the Marine Snail Nerita versicolor Subject: A very powerful proteinaceous inhibitor of metallocarboxypeptidases has been isolated from the marine snail Nerita versicolor and characterized in depth. The most abundant of four, very similar isoforms, NvCla, was taken as reference and N-terminally sequenced to obtain a 372-nucleotide band coding for the protein cDNA. The mature protein contains 53 residues and three disulphide bonds. NvCIa and the other isoforms show an exceptionally high inhibitory capacity of around 1.8 pM for human Carboxypeptidase A1 (hCPA1) and for other A-like members of the M14 CPA subfamily, whereas a twofold decrease in inhibitory potency is observed for carboxypeptidase B-like members as hCPB and hTAFIa. A recombinant form, rNvCI, was produced in high yield and HPLC, mass spectrometry and spectroscopic analyses by CD and NMR indicated its homogeneous, compact and thermally resistant nature. Using antibodies raised with rNvCI and histochemical analyses, a preferential distribution of the inhibitor in the surface regions of the animal body was observed, particularly nearby the open entrance of the shell and gut, suggesting its involvement in biological defense mechanisms. The properties of this strong, small and stable inhibitor of metallocarboxypeptidases envisage potentialities for its direct applicability, as well as leading or minimized forms, in biotechnological/biomedical uses. Keywords: Nerita versicolor; proteinaceous inhibitor; recombinant production; carboxypeptidase; picomolar inhibition; biotechnological and biomedical applications Author: Giovanni Covaleda-Cortés, Martha Hernández, Sebastián Alejandro Trejo, Manuel Mansur, Sergi Rodríguez-Calado, Javier García-Pardo, Julia Lorenzo, Josep Vendrell, María Ángeles Chávez, Maday Alonso-del-Rivero and Francesc Xavier Avilés Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Thu Aug 29 12:22:15 2019 CEST ModDate: Thu Aug 29 12:22:15 2019 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4299275 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SMEUFM+URWPalladioL-Ital Type 1 Custom yes yes no 114 0 BTTJQU+URWPalladioL-Bold Type 1 Custom yes yes no 115 0 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