tailieunhanh - Báo cáo khoa học: Isolation, characterization, sequencing and crystal structure of charybdin, a type 1 ribosome-inactivating protein from Charybdis maritima agg.

A novel, type 1 ribosome-inactivating protein designated charybdin was isolated from bulbs of Charybdis maritimaagg. The protein, consisting of a single polypeptide chain with a molecular mass of 29 kDa, inhibited trans-lation in rabbit reticulocytes with an IC50of nm. Plant genomic DNA extracted from the bulb was amplified by PCR between primers based on the N-terminal and C-terminal sequence of the protein from dissolved crys-tals. | iFEBS Journal Isolation characterization sequencing and crystal structure of charybdin a type 1 ribosome-inactivating protein from Charybdis maritima agg. Eleftherios Touloupakis1 Renate Gessmann2 z Kalliopi Kavelaki1 Emmanuil Christofakis1 Kyriacos Petratos2 and Demetrios F. Ghanotakis1 1 Department of Chemistry University of Crete Greece 2 Institute of Molecular Biology and Biotechnology IMBB FORTH Heraklion Crete Greece Keywords active site Charybdis maritima agg. ribosome-inactivating protein sequence structure Correspondence D. F. Ghanotakis Department of Chemistry University of Crete PO Box 1470 71409 Heraklion Crete Greece Fax 30 2810393601 Tel 30 2810545034 E-mail ghanotakis@ These authors contributed equally to this work Database DNA sequence data from this article have been deposited with the GenBank data library under accession number DQ323742 protein sequence data with UniProt Knowledgebase under accession number P84786 and the crystal structure with the PDB database under accession code 2B7U Received 3 March 2006 revised 18 April 2006 accepted 19 April 2006 A novel type 1 ribosome-inactivating protein designated charybdin was isolated from bulbs of Charybdis maritima agg. The protein consisting of a single polypeptide chain with a molecular mass of 29 kDa inhibited translation in rabbit reticulocytes with an IC50 of nM. Plant genomic DNA extracted from the bulb was amplified by PCR between primers based on the N-terminal and C-terminal sequence of the protein from dissolved crystals. The complete mature protein sequence was derived by partial DNA sequencing and terminal protein sequencing and was confirmed by high-resolution crystal structure analysis. The protein contains Val at position 79 instead of the conserved Tyr residue of the ribosome-inactivating proteins known to date. To our knowledge this is the first observation of a natural substitution of a catalytic residue at the active site of a natural ribosome-inactivating .

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