Isolation and Characterization of a Cone Snail Protease with Homology to CRISP Proteins of the Pathogenesis-related Protein Superfamily Export

@article{Milne2003, abstract = {The pathogenesis-related (PR) protein superfamily is widely distributed in the animal, plant, and fungal kingdoms and is implicated in human brain tumor growth and plant pathogenesis. The precise biological activity of PR proteins, however, has remained elusive. Here we report the characterization, cloning and structural homology modeling of Tex31 from the venom duct of Conus textile. Tex31 was isolated to >95\% purity by activity-guided fractionation using a para-nitroanilide substrate based on the putative cleavage site residues found in the propeptide precursor of conotoxin TxVIA. Tex31 requires four residues including a leucine N-terminal of the cleavage site for efficient substrate processing. The sequence of Tex31 was determined using two degenerate PCR primers designed from N-terminal and tryptic digest Edman sequences. A BLAST search revealed that Tex31 was a member of the PR protein superfamily and most closely related to the CRISP family of mammalian proteins that have a cysteine-rich C-terminal tail. A homology model constructed from two PR proteins revealed that the likely catalytic residues in Tex31 fall within a structurally conserved domain found in PR proteins. Thus, it is possible that other PR proteins may also be substrate-specific proteases. 10.1074/jbc.M304843200}, author = {Milne, Trudy J. and Abbenante, Giovanni and Tyndall, Joel D. and Halliday, Judy and Lewis, Richard J.}, citeulike-article-id = {3269223}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M304843200}, citeulike-linkout-1 = {http://www.jbc.org/cgi/content/abstract/278/33/31105}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/12759345}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=12759345}, day = {15}, doi = {10.1074/jbc.M304843200}, journal = {J. Biol. Chem.}, keywords = {allergens, family, modeling, pathogenesis-related, pr1}, month = {August}, number = {33}, pages = {31105--31110}, posted-at = {2008-09-15 15:13:39}, priority = {0}, title = {Isolation and Characterization of a Cone Snail Protease with Homology to CRISP Proteins of the Pathogenesis-related Protein Superfamily}, url = {http://dx.doi.org/10.1074/jbc.M304843200}, volume = {278}, year = {2003} }

TY - JOUR ID - Milne2003 L3 - citeulike-article-id:3269223 N2 - The pathogenesis-related (PR) protein superfamily is widely distributed in the animal, plant, and fungal kingdoms and is implicated in human brain tumor growth and plant pathogenesis. The precise biological activity of PR proteins, however, has remained elusive. Here we report the characterization, cloning and structural homology modeling of Tex31 from the venom duct of Conus textile. Tex31 was isolated to >95% purity by activity-guided fractionation using a para-nitroanilide substrate based on the putative cleavage site residues found in the propeptide precursor of conotoxin TxVIA. Tex31 requires four residues including a leucine N-terminal of the cleavage site for efficient substrate processing. The sequence of Tex31 was determined using two degenerate PCR primers designed from N-terminal and tryptic digest Edman sequences. A BLAST search revealed that Tex31 was a member of the PR protein superfamily and most closely related to the CRISP family of mammalian proteins that have a cysteine-rich C-terminal tail. A homology model constructed from two PR proteins revealed that the likely catalytic residues in Tex31 fall within a structurally conserved domain found in PR proteins. Thus, it is possible that other PR proteins may also be substrate-specific proteases. 10.1074/jbc.M304843200 IS - 33 JF - J. Biol. Chem. EP - 31110 TI - Isolation and Characterization of a Cone Snail Protease with Homology to CRISP Proteins of the Pathogenesis-related Protein Superfamily VL - 278 SP - 31105 KW - allergens KW - family KW - modeling KW - pathogenesis-related KW - pr1 AU - Milne, Trudy AU - Abbenante, Giovanni AU - Tyndall, Joel AU - Halliday, Judy AU - Lewis, Richard PY - 2003/08/15/ UR - http://dx.doi.org/10.1074/jbc.M304843200 ER -