Isolation and characterization of a neutral phosphatase from wheat seedlings
Thesis/Dissertation
·
OSTI ID:6810390
A neutral phosphatase was purified to homogeneity from wheat seedlings. The enzyme was a monomeric glycoprotein exhibiting a molecular weight of 35,000, frictional ratio of 1.22, Stokes' radius of 26 A, and sedimentation coefficient of 3.2 S. That the enzyme was a glycoprotein was surmised from its chromatographic property on Concanavalin A-Sepharose column. The phosphatase activity was assayed using either fructose-2,6-bisphosphate or p-nitrophenyl phosphate as substrate. The phosphatase activity was not affected by high concentrations of chelating agents and did not require the addition of Mg{sup +2} or Ca{sup +2} for its activity. Molybdate, orthovanadate, Zn{sup +2}, and Hg{sup +2} were all potent inhibitors of the phosphatase activity. The inhibition by Hg{sup +2} was reversed by dithiothreitol. The enzyme activity was stimulated by Mn{sup +2} about 2-fold. On the other hand, 3-phosphoglycerate, fructose-6-P and Pi as well as polyamines inhibited the enzyme activity. The ability of the neutral phosphatase to dephosphorylate protein phosphotyrosine was also investigated. The phosphotyrosyl-substrates, such as ({sup 32}P) phosphotyrosyl-poly(Glu, Tyr)n, -alkylated bovine serum albumin, -angiotensin-1, and -band 3 of erythrocytes, were all substrates of the phosphatase. On the other hand, the enzyme had no activity toward protein phosphoserine and protein phosphothreonine.
- Research Organization:
- Illinois Univ., Chicago, IL (USA)
- OSTI ID:
- 6810390
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CEREALS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYME ACTIVITY
ENZYME INHIBITORS
ENZYMES
ESTERASES
FRACTIONATION
GRASS
HYDROLASES
INHIBITION
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LILIOPSIDA
MAGNOLIOPHYTA
MOLECULAR WEIGHT
NUCLEI
ODD-ODD NUCLEI
PHOSPHATASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLANTS
RADIOISOTOPES
SEEDLINGS
SEPARATION PROCESSES
TRACER TECHNIQUES
WHEAT
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CEREALS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYME ACTIVITY
ENZYME INHIBITORS
ENZYMES
ESTERASES
FRACTIONATION
GRASS
HYDROLASES
INHIBITION
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LILIOPSIDA
MAGNOLIOPHYTA
MOLECULAR WEIGHT
NUCLEI
ODD-ODD NUCLEI
PHOSPHATASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLANTS
RADIOISOTOPES
SEEDLINGS
SEPARATION PROCESSES
TRACER TECHNIQUES
WHEAT