Antibody for detection and quantitation of membrane-associated folate-binding protein from Lactobacillus casei |
| |
| Authors: | L E Pope S A Minskoff C M Chang K S Vitols F M Huennekens |
| |
| Affiliation: | 1. Division of Biochemistry, Department of Basic and Clinical Research, Scripps Clinic and Research Foundation, La Jolla, California 92037 USA;2. Electron Microscopy Laboratory, Scripps Clinic and Research Foundation, La Jolla, California 92037 USA;1. Department of Gastroenterology, Qilu Hospital of Shandong University, Jinan, Shandong, China;2. Laboratory of Translational Gastroenterology, Qilu Hospital of Shandong University, Jinan, Shandong, China;3. Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China;4. Shandong Provincial Clinical Research Center for Digestive Disease, Qilu Hospital of Shandong University, Jinan, Shandong, China;5. Advanced Medical Research Institute, Shandong University, Jinan, China;6. Robot Engineering Laboratory for Precise Diagnosis and Therapy of GI Tumor, Qilu Hospital of Shandong University, Jinan, Shandong, China;1. College of Animal Science and Technology, Zhejiang A & F University, Lin''an, Zhejiang Province, People''s Republic of China;2. College of Animal Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou, People''s Republic of China;3. Department of Biomedical Sciences and Pathobiology, Virginia—Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA;1. Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea;2. iNtRON Biotechnology, Inc., Room 903, JungAng Induspia, 137, Sagimakgol-ro, Jungwon-gu, Seongnam, Gyeonggi-do 13202, Republic of Korea;3. Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea;1. NHC/Ministry of Education/Liaoning Province Key Laboratory of Immunodermatology, National Joint Engineering Research Center for Diagnosis and Treatment of Skin Diseases, The First Hospital of China Medical University, Shenyang, Liaoning Province, 110001, PR China;2. Institute of Respiratory Disease, The First Hospital of China Medical University, Shenyang, Liaoning Province, 110001, PR China;1. Department of Postharvest Science, ARO, The Volcani Center, Rishon LeZion, Israel;2. Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Avda. Agustin Escardino, Paterna, Valencia, Spain;3. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, China;4. Appalachian Fruit Research Station, USDA-ARS, Wiltshire Road, Kearneysville, WV, USA;5. Department of Botany, K.N.P.G. College, Raebareli, India |
| |
| Abstract: | Lactobacillus casei cells contain a 25 kDa, membrane-associated, folate-binding protein (fbp), which is a component of the folate transport system. Polyclonal antibody to fbp (anti-fbp) has been prepared, and conditions have been established for detection and quantitation of the protein. Anti-fbp did not block [3H]folate transport or binding in L. casei cells. As judged by Western blots, the antibody reacted only with fbp on sodium dodecyl sulfate electrophoretograms of Triton X-100 extracts of L. casei membranes. Anti-fbp showed no cross-reactivity with L. casei dihydrofolate reductase, L. casei 5,10-methenyltetrahydrofolate synthetase, L1210 dihydrofolate reductase, rat liver dihydrofolate reductase, or L1210 folate-binding protein. Enzyme-linked immunosorbent assay measurements indicated the presence of an fbp in membranes of Lactobacillus salivarius and two transport-defective sublines of L. casei. Anti-fbp was used to demonstrate selective extraction, with n-butanol, of fbp from a mixture of Triton-solubilized L. casei membrane proteins; repression of fbp in membranes of L. casei cells grown on high levels of folate; and localization of fbp by electron microscopy, using anti-fbp in conjunction with goat anti-rabbit IgG gold conjugate, in L. casei membranes. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
