Purification of Legiobactin and importance of this siderophore in lung infection by Legionella pneumophila. Export

@article{citeulike:5307329, abstract = {When cultured in a low-iron medium, Legionella pneumophila secretes a siderophore (legiobactin) that is both reactive in the chrome azurol S (CAS) assay and capable of stimulating the growth of iron-starved legionellae. Using anion-exchange high-pressure liquid chromatography (HPLC), we purified legiobactin from culture supernatants of a virulent strain of L. pneumophila. In the process, we detected the ferrated form of legiobactin as well as other CAS-reactive substances. Purified legiobactin had a yellow-gold color and absorbed primarily from 220 nm and below. In accordance, nuclear magnetic resonance spectroscopy revealed that legiobactin lacks aromatic carbons, and among the 13 aliphatics present, there were 3 carbonyls. When examined by HPLC, supernatants from L. pneumophila mutants inactivated for lbtA and lbtB completely lacked legiobactin, indicating that the LbtA and LbtB proteins are absolutely required for siderophore activity. Independently derived lbtA mutants, but not a complemented derivative, displayed a reduced ability to infect the lungs of A/J mice after intratracheal inoculation, indicating that legiobactin is required for optimal intrapulmonary survival by L. pneumophila. This defect, however, was not evident when the lbtA mutant and its parental strain were coinoculated into the lung, indicating that legiobactin secreted by the wild type can promote growth of the mutant in trans. Legiobactin mutants grew normally in murine lung macrophages and alveolar epithelial cells, suggesting that legiobactin promotes something other than intracellular infection of resident lung cells. Overall, these data represent the first documentation of a role for siderophore expression in the virulence of L. pneumophila.}, author = {Allard, Kimberly A. and Dao, Jenny and Sanjeevaiah, Prakash and McCoy-Simandle, Kessler and Chatfield, Christa H. and Crumrine, David S. and Castignetti, Domenic and Cianciotto, Nicholas P.}, citeulike-article-id = {5307329}, citeulike-linkout-0 = {http://dx.doi.org/10.1128/IAI.00087-09}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/19398549}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=19398549}, doi = {10.1128/IAI.00087-09}, issn = {1098-5522}, journal = {Infection and immunity}, keywords = {legionella}, month = {July}, number = {7}, pages = {2887--2895}, posted-at = {2009-07-31 09:12:02}, priority = {2}, title = {Purification of Legiobactin and importance of this siderophore in lung infection by Legionella pneumophila.}, url = {http://dx.doi.org/10.1128/IAI.00087-09}, volume = {77}, year = {2009} }

TY - JOUR ID - citeulike:5307329 L3 - citeulike-article-id:5307329 N2 - When cultured in a low-iron medium, Legionella pneumophila secretes a siderophore (legiobactin) that is both reactive in the chrome azurol S (CAS) assay and capable of stimulating the growth of iron-starved legionellae. Using anion-exchange high-pressure liquid chromatography (HPLC), we purified legiobactin from culture supernatants of a virulent strain of L. pneumophila. In the process, we detected the ferrated form of legiobactin as well as other CAS-reactive substances. Purified legiobactin had a yellow-gold color and absorbed primarily from 220 nm and below. In accordance, nuclear magnetic resonance spectroscopy revealed that legiobactin lacks aromatic carbons, and among the 13 aliphatics present, there were 3 carbonyls. When examined by HPLC, supernatants from L. pneumophila mutants inactivated for lbtA and lbtB completely lacked legiobactin, indicating that the LbtA and LbtB proteins are absolutely required for siderophore activity. Independently derived lbtA mutants, but not a complemented derivative, displayed a reduced ability to infect the lungs of A/J mice after intratracheal inoculation, indicating that legiobactin is required for optimal intrapulmonary survival by L. pneumophila. This defect, however, was not evident when the lbtA mutant and its parental strain were coinoculated into the lung, indicating that legiobactin secreted by the wild type can promote growth of the mutant in trans. Legiobactin mutants grew normally in murine lung macrophages and alveolar epithelial cells, suggesting that legiobactin promotes something other than intracellular infection of resident lung cells. Overall, these data represent the first documentation of a role for siderophore expression in the virulence of L. pneumophila. IS - 7 JF - Infection and immunity SN - 1098-5522 EP - 2895 TI - Purification of Legiobactin and importance of this siderophore in lung infection by Legionella pneumophila. VL - 77 SP - 2887 KW - legionella AU - Allard, Kimberly AU - Dao, Jenny AU - Sanjeevaiah, Prakash AU - McCoy-Simandle, Kessler AU - Chatfield, Christa AU - Crumrine, David AU - Castignetti, Domenic AU - Cianciotto, Nicholas PY - 2009/07// UR - http://dx.doi.org/10.1128/IAI.00087-09 ER -