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Regulation of glyoxysomal enzymes during germination of cucumber 3. in vitro translation and characterization of 4 glyoxysomal enzymes


, : Regulation of glyoxysomal enzymes during germination of cucumber 3. in vitro translation and characterization of 4 glyoxysomal enzymes. Plant Physiology (Rockville) 65(1): 40-46

Monospecific antibodies raised against 4 glyoxysomal enzymes (isocitrate lyase, catalase, malate synthase and malate dehydrogenase) were used to detect these proteins among the products of in vitro translation in a wheat germ system programmed with cotyledonary RNA from cucumber seedlings. In vitro immunoprecipitates were compared electrophoretically with the same enzymes labeled in vivo and also with the purified proteins. Isocitrate lyase yields 2 bands on sodium dodecyl sulfate-polyacrylamide gels, as synthesized both in vitro (61.5K and 60K products) and in vivo (63K and 61.5K polypeptides). Both the 63K and 61.5K subunits can also be demonstrated for the isolated enzyme. The 2 subunits are antigenically cross-reactive and yield similar electrophoretic profiles upon partial proteolytic digestion. A larger subunit is seen in vitro than in vivo for both malate dehydrogenase (38K vs. 33K) and catalase (55K vs. 54K); this suggests a need for processing which is often a characteristic of proteins that must be transported across or into membranes. Malate synthase has a MW of 57K in vitro and in vivo, but the isolated enzyme is a glycoprotein, containing N-acetyl glucosamine, mannose, and possibly also fucose and xylose. This indicates that the polypeptide portion of the isolated enzyme is smaller than the in vitro product and suggests processing of malate synthase also. None of the other 3 enzymes appears to be glycosylated. The implications of these size differences for the compartmentalization of matrix and membrane-bound glyoxysomal enzymes are discussed.

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Related references

Riezman, H.; Weir, E.M.; Leaver, C.J.; Titus, D.E.; Becker, W.M., 1980: Regulation of Glyoxysomal Enzymes during Germination of Cucumber: 3. IN VITRO TRANSLATION AND CHARACTERIZATION OF FOUR GLYOXYSOMAL ENZYMES. Monospecific antibodies raised against four glyoxysomal enzymes (isocitrate lyase, catalase, malate synthase, and malate dehydrogenase) have been used to detect these proteins among the products of in vitro translation in a wheat germ system progr...

Becker, W.M.; Leaver, C.J.; Weir, E.M.; Riezman, H., 1978: Regulation of Glyoxysomal Enzymes during Germination of Cucumber: I. Developmental Changes in Cotyledonary Protein, RNA, and Enzyme Activities during Germination. Cytoplasmic rRNA in the cotyledons increased 10-fold between days 0 and 5 of germination, the steepest rise coinciding with the most rapid rise in activities of the two glyoxysomal enzymes isocitrate lyase and malate synthase. Chloroplast rRNA and...

Becker, W.M.; Leaver, C.J.; Weir, E.M.; Riezman, H., 1978: Regulation of glyoxysomal enzymes during germination of cucumber part 1 developmental changes in cotyledonary protein rna and enzyme activities during germination. Developmental patterns of glyoxylate cycle and photosynthetic activities were correlated with electrophoretic profiles of cotyledonary RNA and protein in both light- and dark-grown cucumber seedlings (Cucumis sativus L.). Cytoplasmic rRNA increase...

Lamb, J.E.; Riezman, H.; Becker, W.M., 1978: Regulation of Glyoxysomal Enzymes during Germination of Cucumber: 2. Isolation and Immunological Detection of Isocitrate Lyase and Catalase. The glyoxysomal enzymes isocitrate lyase and catalase have been isolated from etiolated cucumber (Cucumis sativus) cotyledons. The enzymes co-purified through polyethyleneimine precipitation and (NH(4))(2)SO(4) precipitation, and were resolved by...

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Becker, W.; Riezman, H.W.ir, E.; Titus, D.; Leaver, C., 1982: In vitro synthesis and compartmentalization of glyoxysomal enzymes from cucumber. Annals of the New York Academy of Sciences86(386): 329-349

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Koeller, W.; Kindl, H., 1977: Glyoxylate cycle enzymes of the glyoxysomal membrane from cucumber cotyledons. Glyoxysomes were isolated from etiolated cotyledons of cucumber [Cucumis sativus] seedlings. After separation of matrix proteins from the glyoxysomal membranes, enzymes were solubilized from the membranes by 100 mM MgCl2 and purified by sedimentat...