Abstract
InbredDrosophila melanogaster stocks were surveyed for α-glucosidases with nondenaturing gel electrophoresis using a fluorogenic substrate to stain the gels. The glucosidase most active under these conditions is polymorphic. We established that the polymorphism is genetic in origin and that the glucosidase was not likely to be a previously characterized enzyme. The gene encoding the enzyme was mapped cytogenetically to 33 A1-2- 33A8-B1, confirming that this is an enzyme not yet reported inD. melanogaster. The enzyme was partially purified by elution from nondenaturing gels, which enabled us to establish that it has optimal activity at pH 6 and interacts most strongly with α-1–4 glucosides. A developmental and tissue survey suggested that this enzyme could have a purely digestive role or be involved in carbohydrate metabolism inside the organism. We propose that this enzyme is involved in either starch digestion or glycogen metabolism.
Similar content being viewed by others
References
Abraham, I., and Doane, W. W. (1978). Genetic regulation of tissue specific expression ofAmylase structural genes inDrosophila melanogaster.Proc. Natl. Acad. Sci. USA 75(9):4446.
Akam, M. E., Roberts, D. B., and Wolfe, J. (1978).Drosophila haemolymph proteins: Purification, characterization and genetic mapping of larval serum protein 2 inD. melanogaster.Biochem. Genet. 16101.
Ashburner, M. (1989).Drosophila a Laboratory Handbook Cold Spring Harbor Press, Cold Spring Harbor, NY.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72248.
Burns, D. M., and Touster, O. (1982). Purification and characterization of glucosidase II, an endoplasmic reticulum hydrolase involved in glycoprotein biosynthesis.J. Biol. Chem. 257(17):9991.
Burton, R. S., and La Spada, A. (1986). Trehalase polymorphism inDrosophila melanogaster.Biochem. Genet. 24715.
Diem, K., and Lentnor, E. (eds.) (1970).Scientific Tables 7th ed., Ciba-Giegy, Basel, pp. 186–187.
Dowd, J. E., and Riggs, D. S. (1965). A comparison of estimates of Michaelis-Menton kinetic constants from various linear transformations.J. Biol. Chem. 240863.
Faber, C. N., and Glew, R. H. (1984). α-D-Mannosidase. In Bergmeyer, H. U. (ed.),Methods of Enzymatic Analysis, Vol. 4; 3rd ed., Verlag Chemie, Weinheim, pp. 230–240.
Ferreira, C., and Terra, W. R. (1980). Intracellular distribution of hydrolases in midgut caeca cells from an insect with an emphasis on plasma membrane-bound enzymes.Comp. Biochem. Physiol. 66B467.
Frei, E., Schuh, R., Baumgartner, S., Burri, M., Noll, M., Jürgens, G., Seifert, E., Nauber, V., and Jäckle, H., (1988). Molecular characterisation of spalt a homeotic gene required for head and tail development in the Drosophila embryo.EMBO. J. 7197.
Gade, R. (1991). Hyperglycaemia or hypertrehalosaemia? The effect of insect neuropeptides on haemolymph sugars.J. Insect Physiol. 37483.
Hames, B. D. (1981). An introduction to polyacrylamide gels. In Hames, B. D., and Rickwood, D. (eds.),Gel Electrophoresis of Proteins, a Practical Approach, IRL Press.
Herbert, D. N., Foellmer, B., and Helenius, A. (1995). Glucose trimming and reglucosylation determine glycoprotein association with calnexin in the endoplasmic reticulum.Cell 81425.
Hers, H. G. (1963). α-Glucosidase deficiency in generalized glycogen storage disease (Pompe's disease).Biochem. J. 8611.
Hickey, D. A., Benkel, B. F., Abukashawa, S., and Haus, S. (1986). DNA rearrangement causes multiple changes in gene expression at theAmylase locus inDrosophila melanogaster.Biochem. Genet. 26(11/12):757.
Hubbard, S. C., and Ivatt, R. J. (1981). Synthesis and processing of asparagine-linked oligosaccharides.Annu. Rev. Biochem. 50555.
Huber, R. E., and Lefebvre, Y. A. (1971). The purification and some properties of soluble trehalase and sucrase fromDrosophila melanogaster.Can. J. Biochem. 491155.
Jürgens, G. (1988) Head and tail development of theDrosophila embryo involvesspalt, a novel homeotic gene.EMBO J. 7(1):189.
LeJune, N., Thinès-Sempoux, P., and Hers, H. G. (1963). Tissue fractionation studies: Intracellular distribution and properties of α-glucosidases in rat liver.Biochem. J. 8616.
Lindsley, D. L., and Zimm, G. G. (1992).The Genome of Drosophila melanogaster Academic Press, New York.
Mange, A. P., and Sandler, L. (1973). A note on the maternal effect mutantsdaughterless andabnormal oocyte inDrosophila melanogaster.Genetics 7373.
Oliver, M. J., and Williamson, J. H. (1979). Genetic and biochemical aspects of sucrase fromDrosophila melanogaster.Biochem. Genet. 17(9/10):897.
Oliver, M. J., Huber, R. E., and Williamson, J. H. (1978). Genetic and biochemical aspects of trehalase fromDrosophila melanogaster.Biochem. Genet. 16(9/10):927.
Parker, G. F. (1993). D.Phil thesis, Oxford University, Oxford.
Parker, G. F., Williams, P. J., Butters, T. D., and Roberts, D. B. (1991). Detection of the lipid-linked precursor oligosaccharide of N-linked protein glycosylation inDrosophila melanogaster.FEBS Lett. 209(1):58.
Peterson, G. L. (1983). Determination of total protein.Methods Enzymol. 9195.
Roberts, D. B. (ed.) (1986).Drosophila: A Practical Approach, IRL Press.
Roberts, D. B., and Evans-Roberts, S. (1979). The genetic and cytogenetic localization of the three structural genes coding for the major protein ofDrosophila larval serum.Genetics 93663.
Sandler, L. (1977). Evidence for a set of closely linked autosomal genes that interact with sex-chromosome heterochromatin inDrosophila melanogaster.Genetics 86567.
Steele, J. (1982). Glycogen phosphorylase in insects.Insect Biochem. 12(2):131.
Stoll, V. S., and Blanchard, J. S. (1990) Buffers: Principle and practice.Methods Enzymol. 18224.
Tanimura, T., Kitamura, K., Fukuda, T., and Kikuchi, T. (1979). Purification and partial characterisation of three forms of α-glucosidase from the fruit flyDrosophila melanogaster.J. Biochem. 85123.
Terra, W. R., and Ferreira, C. (1981). The physiological role of the peritrophic membrane and trehalase digestive enzymes in the midgut and excreta of starved larvae ofRhynchosciara. J. Insect Physiol.27(5):325.
Terra, W. R., and Jordão, B. (1989). Final digestion of starch inMusca domestica larvae. Distribution and properties of midgut α-D-glucosidases and glucoamylase.Insect Biochem. 19(3):285.
Wigglesworth, V. D. (1972).The Principles of Insect Physiology 7th ed. Chapman and Hall, London.
Wyatt, G. R. (1972). The biochemistry of sugars and polysaccharides in insects.Adv. Insect Physiol. 4287.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Parker, G.F., Roberts, D.B. AGI, a previously unreportedD. melanogaster α-glucosidase: Partial purification, characterization, and cytogenetic mapping. Biochem Genet 34, 117–131 (1996). https://doi.org/10.1007/BF00553608
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00553608