Intracellular Localization of Peptide Hydrolases in Wheat (Triticum aestivum L.) Leaves.

Protoplasts from 8- to 9-day-old wheat (Triticum aestivum L.) leaves were used to isolate organelles which were examined for their contents of peptide hydrolase enzymes and, in the case of vacuoles, other acid hydrolases. High yields of intact chloroplasts were obtained using both equilibrium density gradient centrifugation and velocity sedimentation centrifugation on sucrose-sorbitol gradients. Aminopeptidase activity was found to be distributed, in approximately equal proportions, between the chloroplasts and cytoplasm. Leucyltyrosine dipeptidase was mainly found in the cytoplasm, although about 27% was associated with the chloroplasts. Vacuoles shown to be free from Cellulysin contamination contained all of the protoplast carboxypeptidase and hemoglobin-degrading activities. The acid hydrolases, phosphodiesterase, acid phosphatase, alpha-mannosidase, and beta-N-acetylglucosamidase were found in the vacuole to varying degrees, but no beta-glucosidase was localized in the vacuole.

[1]  V. Wittenbach,et al.  Subcellular localization of proteases in wheat and corn mesophyll protoplasts. , 1981, Plant physiology.

[2]  R. Huffaker,et al.  Hydrolysis of radioactively-labelled ribulose-1,5-bisphosphate carboxylase by an endopeptidase from the primary leaf of barley seedlings , 1981 .

[3]  E. Herman,et al.  Protein bodies of mung bean cotyledons as autophagic organelles. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. J. Dalling,et al.  Proteolytic enzymes in green wheat leaves II. Purification by affinity chromatography, and some properties of proteinases with acid pH optima , 1978 .

[5]  M. Nishimura,et al.  Hydrolases in vacuoles from castor bean endosperm. , 1978, Plant physiology.

[6]  P. Matile.  Biochemistry and Function of Vacuoles , 1978 .

[7]  Murray,et al.  Acid Phosphatase Activities in Developing Seeds of Pisum sativum L , 1977 .

[8]  J. Kruger,et al.  Purification and properties of two proteolytic enzymes with carboxypeptidase activity in germinated wheat. , 1976, Plant physiology.

[9]  C. Ryan.  Proteolytic Enzymes and Their Inhibitors in Plants , 1973 .

[10]  C. Schnarrenberger,et al.  Development of Microbodies in Sunflower Cotyledons and Castor Bean Endosperm during Germination. , 1971, Plant physiology.

[11]  J. Mikola,et al.  Partial purification and enzymatic properties of an aminopeptidase from barley. , 1971, Archives of biochemistry and biophysics.

[12]  E. Noltmann,et al.  [23] Glucose 6-phosphate dehydrogenase (crystalline) from brewers' yeast , 1966 .

[13]  M. Gibbs,et al.  Enzymes of Glycolysis , 1964 .

[14]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[15]  D. Arnon.  COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. , 1949, Plant physiology.

[16]  W. Crocker.  Catalase , 1911, Botanical Gazette.