Biochemical characterization of the plasma membrane H/sup +/ - ATPase from red beet (Beta vulgaris) hypocotyl tissue
Thesis/Dissertation
·
OSTI ID:6592051
Several biochemical techniques including selective solubilization followed by gel filtration or various types of affinity chromatography, and antibody production were employed in an attempt to purify the plasma membrane H/sup +/ - ATPase from red beet hypocotyl tissue. While the enzyme could not be purified using any of these methods, it was possible to successfully conduct a more detailed biochemical analysis of the H/sup +/ - ATPase. The molecular weight and isoelectric point of the enzyme were determined using N,N'dicyclohexylcarbodiimide (DCCD) and a H/sup +/ - ATPase antibody. When plasma membrane vesicles were incubated with 20 ..mu..M (/sup 14/C)-DCCD at 0 C, a single 97,000 dalton protein was apparent on a fluorograph. A close correlation between (/sup 14/C)-DCCD labelling of the 97,000 dalton protein and the extent of ATPase inhibition over a range of DCCD concentrations suggests that this 97,000 dalton protein is a component of the plasma membrane H/sup +/ - ATPase. An antibody raised against the plasma membrane H/sup +/ - ATPase of Neurospora crassa cross-reacted with the 97,000 dalton DCCD-binding protein, further supporting the identity of this protein. Immunoblots of two dimensional gels of red beet plasma membrane vesicles indicated the isoelectric point of the enzyme to be pH 6.5.
- Research Organization:
- California Univ., Davis (USA)
- OSTI ID:
- 6592051
- Country of Publication:
- United States
- Language:
- English
Similar Records
H/sup +/-ATPase activity from storage tissue of Beta vulgaris. IV. N,N'-dicyclohexylcarbodiimide binding and inhibition of the plasma membrane H/sup +/-ATPase
Inhibition of the coated vesicle proton pump and labeling of a 17,000-dalton polypeptide by N,N'-dicyclohexylcarbodiimide
Size of the plasma membrane H -ATPase from Neurospora crassa determined by radiation inactivation and comparison with the sarcoplasmic reticulum CaS -ATPase from skeletal muscle
Journal Article
·
Sat Feb 28 23:00:00 EST 1987
· Plant Physiol.; (United States)
·
OSTI ID:6550876
Inhibition of the coated vesicle proton pump and labeling of a 17,000-dalton polypeptide by N,N'-dicyclohexylcarbodiimide
Journal Article
·
Sat Aug 15 00:00:00 EDT 1987
· J. Biol. Chem.; (United States)
·
OSTI ID:6375590
Size of the plasma membrane H -ATPase from Neurospora crassa determined by radiation inactivation and comparison with the sarcoplasmic reticulum CaS -ATPase from skeletal muscle
Journal Article
·
Thu Jul 25 00:00:00 EDT 1985
· J. Biol. Chem.; (United States)
·
OSTI ID:6179167
Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
ATP-ASE
BEETS
CARBON 14 COMPOUNDS
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
ELECTROPHORESIS
ENZYMES
FOOD
FRACTIONATION
HYDROLASES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MEMBRANES
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PHOSPHOHYDROLASES
PLANTS
PROTEINS
SEPARATION PROCESSES
TRACER TECHNIQUES
VEGETABLES
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
ATP-ASE
BEETS
CARBON 14 COMPOUNDS
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
ELECTROPHORESIS
ENZYMES
FOOD
FRACTIONATION
HYDROLASES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MEMBRANES
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PHOSPHOHYDROLASES
PLANTS
PROTEINS
SEPARATION PROCESSES
TRACER TECHNIQUES
VEGETABLES