Assay of ribulose bisphosphate carboxylase

Conference · Wed Apr 01 00:00:00 EST 1987 · Plant Physiol., Suppl.; (United States)

OSTI ID:5878370

Pike, C; Berry, J

Assays of ribulose bisphosphate carboxylase (rubisco) can be used to illustrate many properties of photosynthetic systems. Many different leaves have been assayed with this standard procedure. The tissue is ground with a mortar and pestle in extraction buffer. The supernatant after centrifugation is used as the source of enzyme. Buffer, RuBP, (/sup 14/C)-NaHCO/sub 3/, and enzyme are combined in a scintillation vial; the reaction is run for 1 min at 30/sup 0/. The acid-stable products are counted. Reproducibility in student experiments has been excellent. The assay data can be combined with analyses of leaf properties such as fresh and dry weight, chlorophyll and protein content, etc. Students have done projects such as the response of enzyme to temperature and to various inhibitors. They also report on the use of a transition state analog, carboxyarabinitol bisphosphate, to titrate the molar concentration of rubisco molecules (active sites) in an enzyme sample. Thus, using crude extracts the catalytic activity of a sample can be compared to the absolute quantity of enzyme or to the turnover number.

• Research Organization: Franklin and Marshall College, Lancaster, PA
• OSTI ID: 5878370
• Report Number(s): CONF-8707108-; TRN: 87-039111
• Journal Information: Plant Physiol., Suppl.; (United States), Vol. 83:4; Conference: Annual meeting of the American Society of Plant Physiologists, St. Louis, MO, USA, 19 Jul 1987
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CARBOXYLASE
ENZYME ACTIVITY
QUANTITATIVE CHEMICAL ANALYSIS
CARBON 14 COMPOUNDS
CENTRIFUGATION
SCINTILLATION COUNTING
TRACER TECHNIQUES
CARBON-CARBON LYASES
CARBOXY-LYASES
CHEMICAL ANALYSIS
COUNTING TECHNIQUES
ENZYMES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LYASES
SEPARATION PROCESSES
550201* - Biochemistry- Tracer Techniques