Solubilization of brush borders of hamster small intestine and fractionation of some of the components |
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Authors: | DR Critchley KE Howell A Eichholz |
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Institution: | 1. Department of Biochemistry, The University, Leicester U.K.;2. Yale University, New Haven, Conn. U.S.A.;3. Department of Physiology, College of Medicine and Dentistry of New Jersey, Rutgers Medical School, New Brunswick, N.J. 08903 U.S.A. |
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Abstract: | About 90% of the protein of hamster intestinal brush borders was solubilised in 0.25% (w/v) sodium dodecyl sulphate without total loss of biological activity. Detergent-polyacrylamide gel electrophoresis of the solubilised protein separated 10–15 bands and partially resolved maltase, lactase, sucrase-maltase, trehalase and alkaline phosphatase activities. The disaccharidases, which were associated with the higher molecular weight proteins, were preferentially solubilised with 0.1%. (w/v) Triton X-100, butanol or papain, whereas Tris and NaI extracted only the lower molecular weight proteins, possible derived from the core filaments.Electrophoresis of brush border proteins metabolically labelled with 14C] glucosamine suggested that many of the membrane-bound enzymes are glycoproteins. However, chromatography of a papain digest on Sephadex G-200 showed that the sucrase-maltase complex can be separated nearly free of carbohydrate without total loss of activity.The importance of characterizing membrane proteins solubilised by a number of techniques is discussed. |
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Keywords: | To whom reprint request should be addressed |
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