Partial characterization of chorionic gonadotropin-like binding sites from the bacteria Xanthomonas maltophilia.
The gram-negative bacterium, Xanthomonas maltophilia, has low- and high-affinity luteinizing hormone/chorionic gonadotropin (LH/CG)-binding sites, similar to the LH/CG receptor found in mammals. Although the low-affinity site binds both LH and human CG (hCG), the high-affinity site is specific for hCG. In the current investigation, these two binding sites were independently isolated from X. maltophilia for further characterization. To isolate functional binding sites, we developed a solubilization method using the detergent zwittergent 3,14 and high glycerol concentrations that allowed for the maintenance of ligand-binding integrity. Gel filtration experiments established molecular weights of 170 and 11.5 kDa for the two binding sites, which were supported by data from photoaffinity labeling and ultracentrifugation experiments. Gel filtration data also suggested the presence of a third binding site of 5.4 kDa. The 170-kDa site had a binding affinity of Kd = 12 x 10(-6) and bound both LH and hCG. The small molecular weight site had an affinity of Kd = 9.4 x 10(-8) and was CG specific. Collectively, these data demonstrate the presence of multiple hormone binding sites in X. maltophilia that differ in molecular size, binding affinity, and ligand specificity.
Edwards
Jeffrey G
JG
Department of Physiology, University of Utah School of Medicine, Salt Lake City, Utah 84108-1297, USA. jeffrey_edwards@brown.edu
Odell
William D
WD
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D002850:Chromatography, Gel; D006801:Humans; D007457:Iodine Radioisotopes; D007986:Luteinizing Hormone / Q000378:metabolism; D012995:Solubility; D020615:Stenotrophomonas maltophilia / Q000737:chemistry* / Q000378:metabolism; D014461:Ultracentrifugation
D002352:Carrier Proteins; D006063:Chorionic Gonadotropin; D007457:Iodine Radioisotopes; D007986:Luteinizing Hormone