Abstract
The lipase gene family comprises three vertebrate genes, lipoprotein lipase (LPL), hepatic lipase (HL) and pancreatic lipase (PL), that are derived from a common ancestral gene. While these lipases are functionally related, considerable evidence indicates that LPL and HL share a higher degree of structural homology than either shares with PL (1–4). For example, it has been established by a number of different methods that LPL and HL are functionally active as homodimers, while PL is active as a monomer (5–9). However, there are discrepancies reported in the literature, namely the findings of monomeric functional units of LPL and HL by Ikeda et al. (10) and Schoonderwoerd et al. (11), the latter concluding that the functional unit of rat HL in the liver is a monomer, while in adrenal gland and ovary it might be a dimer.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kirchgessner, T. G., Chuat, J.-C., Heinzmann, C., Etienne, J., Guilhot, S., Svenson, K., Ameis, D., Pilon, C., d’Auriol, L., Andalibi, A., Schotz, M. C., Galibert, F., and Lusis, A. J. (1989) Organization of the human lipoprotein lipase gene and evolution of the lipase gene family. Proc. Natl. Acad. Sci. 86, 9647–9651.
Derewenda, Z. S., and Cambillau, C. (1991) Effects of gene mutations in lipoprotein lipase as interpreted by a molecular model of pancreatic lipase. J. Biol. Chem. 266, 23,112–23,119.
Hide, W. A., Chan, L., and Li, W.-H. (1992) Structure and evolution of the lipase superfamily. J. Lipid Res. 33, 167–178.
van Tilbeurgh, H., Roussel, A., Lalouel, J.-M., and Cambillau, C. (1994) Lipoprotein lipase: molecular model based on the pancreatic lipase X-ray structure: consequences for heparin binding and catalysis. J. Biol. Chem. 269, 4626–4633.
Garfinkel, A. S., Kempner, E. S., Ben-Zeev, O., Nikazy, J., James, S. J., and Schotz, M. C. (1983) Lipoprotein lipase: Size of the functional unit determined by radiation inactivation. J. Lipid Res. 24, 775–780.
Osborne, J. C., Jr., Bengtsson-Olivecrona, G., Lee, N. S., and Olivecrona, T. (1985) Studies on inactivation of lipoprotein lipase: role of the dimer to monomer dissociation. Biochemistry. 24, 5606–5611.
Olivecrona, T., Bengtsson-Olivecrona, G., Osborne, J. C, Jr., and Kempner, E. S. (1985) Molecular size of bovine lipoprotein lipase as determined by radiation inactivation. J. Biol. Chem. 280, 6888–6891.
Hill, J. S., Davis, R. C., Yang, D., Wen, J., Philo, J. S., Poon, P. H., Phillips, M. L., Kempner, E. S., and Wong, H. (1996) Human hepatic lipase subunit structure determination. J. Biol. Chem. 271, 22,931–22,936.
Rovery, M., Boudouard, M., and Bianchietta, J. (1978) An improved large scale procedure for the purification of porcine pancreatic lipase. Biochim. Biophys. Acta. 525, 373–379.
Ikeda, Y., Takagi, A., and Yamamoto, A. (1989) Purification and characterization of lipoprotein lipase and hepatic triglyceride lipase from human postheparin plasma; production of monospecific antibody to the individual lipase. Biochim. Biophys. Acta. 1003, 254–269.
Schoonderwoerd, K., Hom, M. L., Luthjens, L. H., Vieira van Bruggen, D., and Jansen, H. (1996) Functional molecular mass of rat hepatic lipase in liver, adrenal gland and ovary is different. Biochem. J. 318, 463–467.
Vannier, C., and Ailhaud, G. (1989) Biosynthesis of lipoprotein lipase in cultured mouse adypocytes II. Processing, subunit assembly, and intracellular transport. J. Biol. Chem. 264, 13,206–13,216.
Ben-Zeev, O., Doolittle, M. H., Davis, R. C., Elovson, J., and Schotz, M. C. (1992) Maturation of liporotein lipase: expression of full catalytic activity requires glucose trimming but not translocation to the cis-Golgi compartment. J. Biol. Chem. 267, 6219–6227.
Carroll, R., Ben-Zeev, O., Doolittle, M. H., and Severson, D. L. (1992) Activation of lipoprotein lipase in cardiac muscle by glycosylation requires trimming of glucose residues in the endoplasmic reticulum. Biochem. J. 285, 639–696.
Park, J.-W., Oh, M.-S., Yang, J.-Y., Park, B.-H., Rho, H.-W., Lim, S.-N., Jhee, E.-C., and Kim, H.-R. (1995) Glycosylation, dimerization, and heparin affinity of lipoprotein lipase in 3T3-L1 adipocytes. Biochim. Biophys. Acta. 1254, 45–50.
Vannier, C., Deslex, S., Pradines-Figueres, A., and Ailhaud, G. (1989) Biosynthesis of lipoprotein lipase in cultured mouse adipocytes I: Characterization of a specific antibody and relationships between the intracellular and secreted pools of the enzyme. J. Biol. Chem. 264, 13,199–13,205.
Doolittle, M. H., Ben-Zeev, O., Elovson, J., Martin, D., Kirchgessner, T., and Schotz, M. C. (1990) Post-translational regulation of lipoprotein lipase during feeding and fasting. J. Biol. Chem. 265, 4570–4577.
Davis, R. C., Ben-Zeev, O., Martin, D., and Doolittle, M. H. (1990) Combined lipase deficiency in the mouse: lipase transcription, translation and processing. J. Biol. Chem. 265, 17,960–17,966.
Bergö, M., Olivecrona, G., and Olivecrona, T. (1996) Forms of lipoprotein lipase in rat tissues: in adipose tissue the proportion of inactive lipase increases on fasting. Biochem. J. 313, 893–898.
Buscà, R., Martinez, M., Vilella, E., Pognonec, P., Deeb, S., Auwerx, J., Reina, M., and Vilaró, S. (1996) The mutation Gly142-Glu in human lipoprotein lipase produces a missorted protein that is diverted to the lysosomes. J. Biol. Chem. 271, 2139–2146.
Sheeler, P. (1981) Centifugation in Biology and Medical Science. John Wiley and Sons, New York.
Hinton, R., and Dobrota, M. (1976) Density Gradient Centrifugation. North-Holland Publishing Company, Amsterdam.
Price, C. A. (1982) Centrifugation in Density Gradients. Academic Press, New York, London.
Ailhaud, G. (1990) Cellular and secreted lipoprotein lipase revisited. Clin. Biochem. 23, 343–347.
Olive, C., and Levy, H. R. (1975) Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Methods Enzymol. 41, 196–201.
Johnson, H. S., and Hatch, M. D. (1970) Properties and regulation of leaf nicotinamide-adenine dinucleotide phosphate-malate dehydrogenase and “malic” enzyme in plants with the C14-dicarboxylic acid pathway of photosynthesis. Biochem. J. 119, 273–280.
Lowry, O. H., Rosenbrough, N. J., Fair, A. L., and Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.
Wion, K. L., Kirchgessner, T. G., Lusis, A. J., Schotz, M. C., and Lawn, R. M. (1987) Human lipoprotein lipase complementary DNA sequence. Science. 235, 1638–1641.
Ameis, D., Stahnke, G., Kobayashi, J., McLean, J., Lee, G., Büscher, M., Schotz, M. C., and Will, H. (1990) Isolation and characterization of the human hepatic lipase gene. J. Biol. Chem. 265, 6652–6555.
Lowe, M. E., Rosenblum, J. L., and Strauss, A. W. (1989) Cloning and characterization of human pancreatic lipase cDNA. J. Biol. Chem. 264, 20,042–20,048.
Ben-Zeev, O., Stahnke, G., Liu, G., Davis, R. C., and Doolittle, M. H. (1994) Lipoprotein lipase and hepatic lipase: the role of asparagine-linked glycosylation in the expression of a functional enzyme. J. Lipid Res. 35, 1511–1523.
Canalias, F., Visvikis, A., Thioudellet, C., and Siest, G. (1994) Stable expression of enzymatically active human pancreatic lipase in V79 cells: purification and characterization of the recombinant enzyme. Clin. Chem. 40, 1251–1257.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc, Totowa, NJ
About this protocol
Cite this protocol
Ben-Zeev, O., Doolittle, M.H. (1999). Determining Lipase Subunit Structure by Sucrose Gradient Centrifugation. In: Doolittle, M., Reue, K. (eds) Lipase and Phospholipase Protocols. Methods in Molecular Biology™, vol 109. Humana Press. https://doi.org/10.1385/1-59259-581-2:257
Download citation
DOI: https://doi.org/10.1385/1-59259-581-2:257
Publisher Name: Humana Press
Print ISBN: 978-0-89603-546-1
Online ISBN: 978-1-59259-581-5
eBook Packages: Springer Protocols