Characterization and Purification of the Chorionic Gonadotropin-Like Protein Binding Site InCandida Albicans
| Autor: | Omar Caticha, William D. Odell |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
endocrine system
Blotting Western Plasma protein binding Biology Binding Competitive Chorionic Gonadotropin Chromatography Affinity Fungal Proteins Iodine Radioisotopes Endocrinology Xanthomonas Affinity chromatography Western blot Candida albicans medicine Binding site Fungal protein Binding Sites medicine.diagnostic_test Temperature General Medicine Hydrogen-Ion Concentration Ligand (biochemistry) biology.organism_classification Molecular biology Molecular Weight Kinetics Biochemistry Electrophoresis Polyacrylamide Gel Carrier Proteins |
| Zdroj: | Endocrine Research. 20:1-19 |
| ISSN: | 1532-4206 0743-5800 |
| DOI: | 10.1080/07435809409035852 |
| Popis: | Recent studies from our laboratory have shown that human chorionic gonadotropin (hCG), human luteinizing hormone (hLH), and CG-like proteins from Xanthomonas maltophilia (xCG) and Candida albicans (CaCGLP) induce Candida albicans transition from blastospores to hyphal forms. Xanthomonas maltophilia CG-like protein (xCG), hCG, and hLH also bind to Candida albicans blastospores with a high-affinity nM Kd, indicating that these substances can control Candida albicans pathogenicity. The work reported herein describes the purification of the binding site for these CG-like proteins from Candida albicans. The purification developed involved alcohol extraction followed by affinity chromatography. The product obtained was a protein of 64-69 kDa, as analyzed by sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE), Western blot and Sephadex G-100 column chromatography. This binding site reacted in a Western blot with both 125I-hCG and 125I-CaCGLP. Purified CaCGLP was able to displace specifically 125I-hCG bound to Candida albicans blastospores. Scatchard plot analysis showed that the Kd of this reaction was of high affinity in the nM range, and also indicated the presence of one single binding site. These results lead us to conclude: 1) Candida albicans possesses a binding site which is able to bind hCG, hLH and CG-like proteins from Xanthomonas maltophilia and Candida albicans; 2) This binding site is a hydrophobic protein of approximately 64 kDa and; 3) We postulate that CaCGLP is the natural ligand for this binding site and this system is used to control Candida albicans transition and, therefore, pathogenicity. |
| Databáze: | OpenAIRE |
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