| First Author | Meheus LA | Year | 1993 |
| Journal | J Immunol | Volume | 151 |
| Issue | 3 | Pages | 1535-47 |
| PubMed ID | 8335946 | Mgi Jnum | J:17362 |
| Mgi Id | MGI:65409 | Doi | 10.4049/jimmunol.151.3.1535 |
| Citation | Meheus LA, et al. (1993) Identification by microsequencing of lipopolysaccharide-induced proteins secreted by mouse macrophages. J Immunol 151(3):1535-47 |
| abstractText | The conditioned medium of the murine macrophage PU5.1.8 was analyzed by two-dimensional gel electrophoresis in order to detect LPS-induced proteins. Spots of interest were identified by microsequencing of internal peptides generated by limited in situ acid hydrolysis. In total conditioned medium, several monokines (TNF-alpha and macrophage inflammatory protein-1 alpha and 1 beta) were identified as LPS-induced spots. Because minor spots could be masked by the complexity of the 2-D pattern, conditioned medium was successively fractionated by zinc precipitation and affinity chromatography (Procion red and Con A agarose). Zinc supernatant fraction, Procion red flow-through, and Con A eluate fractions were further analyzed by 2-D gel electrophoresis for the presence of LPS-induced spots. In these fractions serum amyloid A3, lipocalin 24p3, cathepsin B, and plasminogen activator inhibitor-I were characterized as LPS-induced proteins secreted by macrophages. Lipocalin 24p3 protein was retrieved for the first time. In addition to these proteins that follow a classical secretory pathway, several cellular proteins (mainly ribosomal proteins) were retrieved as LPS-induced proteins in the conditioned medium. Control experiments argue against the obvious explanation that the latter observation is caused solely by cellular leakage. |
