Abstract
Both transport function and microvillus membrane physical properties evolve as the enterocyte matures and migrates up the crypt-villus axis. We isolated enriched fractions of villus tip, mid-villus, and crypt enterocytes from which microvillus membrane vesicles were prepared. Using this material we characterized the alterations that occur in microvillus membrane fluidity as the rabbit enterocyte matures and correlated these with kinetic studies of glucose transport. With increasing maturity the microvillus membrane becomes more rigid due to both an increase in the cholesterol/phospholipid ratio and alterations in individual phospholipid subclasses. Maximal rates of glucose transport were greatest in microvillus membrane vesicles prepared from mature cells. However, the glucose concentration producing half-maximal rates of transport (Km) was significantly lower in crypt microvillus membrane vesicles, suggesting that a distinct glucose transporter existed in crypt enterocytes. This distinction disappeared when differences between membrane lipid environments were removed. By fluidizing villus-tip microvillus membrane vesicles, in vitro, to levels seen in the crypt microvillus membrane, we observed a reduction in the Km of this transport system. These data suggest that the kinetic characteristics of the sodium-dependent glucose transporter are dependent upon its local membrane environment.
Full text
PDF








Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Brasitus T. A., Dahiya R., Dudeja P. K., Bissonnette B. M. Cholesterol modulates alkaline phosphatase activity of rat intestinal microvillus membranes. J Biol Chem. 1988 Jun 25;263(18):8592–8597. [PubMed] [Google Scholar]
- Brasitus T. A., Dudeja P. K. Alterations in the physical state and composition of brush border membrane lipids of rat enterocytes during differentiation. Arch Biochem Biophys. 1985 Jul;240(1):483–488. doi: 10.1016/0003-9861(85)90054-2. [DOI] [PubMed] [Google Scholar]
- DAHLQVIST A. METHOD FOR ASSAY OF INTESTINAL DISACCHARIDASES. Anal Biochem. 1964 Jan;7:18–25. doi: 10.1016/0003-2697(64)90115-0. [DOI] [PubMed] [Google Scholar]
- FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
- Fine J. B., Sprecher H. Unidimensional thin-layer chromatography of phospholipids on boric acid-impregnated plates. J Lipid Res. 1982 May;23(4):660–663. [PubMed] [Google Scholar]
- Keljo D. J., MacLeod R. J., Perdue M. H., Butler D. G., Hamilton J. R. D-Glucose transport in piglet jejunal brush-border membranes: insights from a disease model. Am J Physiol. 1985 Dec;249(6 Pt 1):G751–G760. doi: 10.1152/ajpgi.1985.249.6.G751. [DOI] [PubMed] [Google Scholar]
- Kessler M., Acuto O., Storelli C., Murer H., Müller M., Semenza G. A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of D-glucose and choline transport systems. Biochim Biophys Acta. 1978 Jan 4;506(1):136–154. doi: 10.1016/0005-2736(78)90440-6. [DOI] [PubMed] [Google Scholar]
- Klemperer H. G., Haynes G. R. Thymidine kinase in rat liver during development. Biochem J. 1968 Jul;108(4):541–546. doi: 10.1042/bj1080541. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knickelbein R. G., Aronson P. S., Dobbins J. W. Membrane distribution of sodium-hydrogen and chloride-bicarbonate exchangers in crypt and villus cell membranes from rabbit ileum. J Clin Invest. 1988 Dec;82(6):2158–2163. doi: 10.1172/JCI113838. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lepage G., Roy C. C. Direct transesterification of all classes of lipids in a one-step reaction. J Lipid Res. 1986 Jan;27(1):114–120. [PubMed] [Google Scholar]
- Meddings J. B. Lipid permeability of rat jejunum and ileum: correlation with physical properties of the microvillus membrane. Biochim Biophys Acta. 1988 Aug 18;943(2):305–314. doi: 10.1016/0005-2736(88)90562-7. [DOI] [PubMed] [Google Scholar]
- Meddings J. B., Scott R. B., Fick G. H. Analysis and comparison of sigmoidal curves: application to dose-response data. Am J Physiol. 1989 Dec;257(6 Pt 1):G982–G989. doi: 10.1152/ajpgi.1989.257.6.G982. [DOI] [PubMed] [Google Scholar]
- Meddings J. B., Theisen S. Development of rat jejunum: lipid permeability, physical properties, and chemical composition. Am J Physiol. 1989 May;256(5 Pt 1):G931–G940. doi: 10.1152/ajpgi.1989.256.5.G931. [DOI] [PubMed] [Google Scholar]
- Melchior D. L., Czech M. P. Sensitivity of the adipocyte D-glucose transport system to membrane fluidity in reconstituted vesicles. J Biol Chem. 1979 Sep 25;254(18):8744–8747. [PubMed] [Google Scholar]
- Molitoris B. A., Kinne R. Ischemia induces surface membrane dysfunction. Mechanism of altered Na+-dependent glucose transport. J Clin Invest. 1987 Sep;80(3):647–654. doi: 10.1172/JCI113117. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Molitoris B. A. Membrane fluidity: measurement and relationship to solute transport. Semin Nephrol. 1987 Mar;7(1):61–71. [PubMed] [Google Scholar]
- Peerce B. E., Wright E. M. Sodium-induced conformational changes in the glucose transporter of intestinal brush borders. J Biol Chem. 1984 Nov 25;259(22):14105–14112. [PubMed] [Google Scholar]
- Pilch P. F., Thompson P. A., Czech M. P. Coordinate modulation of D-glucose transport activity and bilayer fluidity in plasma membranes derived from control and insulin-treated adipocytes. Proc Natl Acad Sci U S A. 1980 Feb;77(2):915–918. doi: 10.1073/pnas.77.2.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rood R. P., Emmer E., Wesolek J., McCullen J., Husain Z., Cohen M. E., Braithwaite R. S., Murer H., Sharp G. W., Donowitz M. Regulation of the rabbit ileal brush-border Na+/H+ exchanger by an ATP-requiring Ca++/calmodulin-mediated process. J Clin Invest. 1988 Sep;82(3):1091–1097. doi: 10.1172/JCI113665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rouser G., Fkeischer S., Yamamoto A. Two dimensional then layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots. Lipids. 1970 May;5(5):494–496. doi: 10.1007/BF02531316. [DOI] [PubMed] [Google Scholar]
- Rowling P. J., Sepúlveda F. V. The distribution of (Na+ + K+)-ATPase along the villus crypt-axis in the rabbit small intestine. Biochim Biophys Acta. 1984 Mar 28;771(1):35–41. doi: 10.1016/0005-2736(84)90107-x. [DOI] [PubMed] [Google Scholar]
- Schachter D., Shinitzky M. Fluorescence polarization studies of rat intestinal microvillus membranes. J Clin Invest. 1977 Mar;59(3):536–548. doi: 10.1172/JCI108669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thulborn K. R., Sawyer W. H. Properties and the locations of a set of fluorescent probes sensitive to the fluidity gradient of the lipid bilayer. Biochim Biophys Acta. 1978 Aug 4;511(2):125–140. doi: 10.1016/0005-2736(78)90308-5. [DOI] [PubMed] [Google Scholar]
- Van Blitterswijk W. J., Van Hoeven R. P., Van der Meer B. W. Lipid structural order parameters (reciprocal of fluidity) in biomembranes derived from steady-state fluorescence polarization measurements. Biochim Biophys Acta. 1981 Jun 22;644(2):323–332. doi: 10.1016/0005-2736(81)90390-4. [DOI] [PubMed] [Google Scholar]
- Yuli I., Wilbrandt W., Shinitzky M. Glucose transport through cell membranes of modified lipid fluidity. Biochemistry. 1981 Jul 21;20(15):4250–4256. doi: 10.1021/bi00518a003. [DOI] [PubMed] [Google Scholar]
- van Blitterswijk W. J., van der Meer B. W., Hilkmann H. Quantitative contributions of cholesterol and the individual classes of phospholipids and their degree of fatty acyl (un)saturation to membrane fluidity measured by fluorescence polarization. Biochemistry. 1987 Mar 24;26(6):1746–1756. doi: 10.1021/bi00380a038. [DOI] [PubMed] [Google Scholar]

