Biosynthesis of polyuronides inMucor rouxii: Partial characterization of fucosyl transferase |
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| Institution: | 1. Instituto de Investigacio´n en Biologia Experimental, Facultad de Quimica, Universidad de Guanajuato, Apartado postal 187, Guanajuato, Gto. C.P. 36000, Me´xico;2. Departamento de Gene´tica y Biologia Molecular, Cinvestav, IPN, Apartado postal 14-740, Me´xico, D.F. CP 07000, Me´xico;1. Department of Viral Therapeutics, Virology Division, United States Army Medical Research Institute of Infectious Disease, Ft. Detrick, Frederick, MD 21702, United States;2. Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick National Laboratory for Cancer Research, Frederick, MD 21702, United States;3. Basic Science Program, Leidos Biomedical Research Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, United States;4. Office of the Chief Scientist, Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, Frederick, MD 2170, United States;5. Division of Clinical Research, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD 20892, United States;1. Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, col. Noria Alta, C.P. 36050 Guanajuato, Gto., Mexico;2. Centro de Investigaciones y de Estudios Avanzados del IPN, Apartado Postal 629, 36500 Irapuato, Guanajuato, Mexico;3. Laboratorio de Glicobiología Humana y Diagnóstico Molecular, Centro de Investigación en Dinámica Celular, Instituto de Investigación en Ciencias Básicas y Aplicada, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, C.P. 62209 Cuernavaca, Mor., Mexico;1. School of Life Science and National Glycoengineering Research Center, Shandong University, Jinan 250100, China;2. Institute of Cardiovascular Disease, General Hospital of Jinan Military Region, Jinan 250022, China |
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| Abstract: | Most of the fucosyl transferase activity fromMucor rouxii was detected in a crude membrane fraction. The enzyme transferredl-fucose from GDP-fucose to endogenous and exogenous acceptors. When crude membrane fractions were treated with neutral detergents such as Trition X-100 or Brij 36 T enzyme activity became dependent on exogenous acceptors such as mucoric acid or mucoran. Brij-treated membrane fractions showed maximum fucosyl transferase activity at pH 6.5, and at a temperature between 22 and 28°C. The cations Mn2+, Mg2+, Co2+, Zn2+, Fe2+, and Ca2+ activated the enzyme about twofold. The former was slightly more stimulatory at 4 mM. Km for GDP-fucose was 10 μM. Evidence was obtained that mucoric acid serves as acceptor for fucosyl moieties. Acid hydrolysis of the product synthesized from GDP-fuc by Brij-treated membrane fractions revealed fucose as the major radioactive sugar. |
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