Active site histidine in spinach ribulosebisphosphate carboxylase/oxygenase modified by diethyl pyrocarbonate
(TH) Diethyl pyrocarbonate was synthesized from (TH) ethanol prepared by the reduction of acetaldehyde by NaB3H4. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) from spinach was inactivated with this reagent at pH 7.0 the presence of 20 mM MgS , and tryptic peptides that contained modified histidine residues were isolated by reverse-phase high-performance liquid chromatography. Labeling of the enzyme was conducted in the presence and absence of the competitive inhibitor sedoheptulose 1,7-bisphosphate. The amount of one peptide that was heavily labeled in the absence of this compound was reduced 10-fold in its presence. The labeled residue was histidine-298. This result, in combination with earlier experiments, suggests that His-298 in spinach RuBisCO is located in the active site domain and is essential to enzyme activity. This region of the primary structure is strongly conserved in seven other ribulosebisphosphate carboxylases from divergent sources.
- Research Organization:
- Washington State Univ., Pullman
- OSTI ID:
- 6259022
- Journal Information:
- Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 15; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AMINO ACIDS
AZOLES
BIOSYNTHESIS
CARBON COMPOUNDS
CARBON-CARBON LYASES
CARBONATES
CARBOXY-LYASES
CARBOXYLASE
CARBOXYLIC ACIDS
ENZYMES
FOOD
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
HISTIDINE
HYDROLASES
IMIDAZOLES
KINETICS
LABELLED COMPOUNDS
LYASES
MOLECULAR STRUCTURE
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXYGEN COMPOUNDS
PEPTIDE HYDROLASES
PLANTS
SERINE PROTEINASES
SPINACH
SYNTHESIS
TRITIUM COMPOUNDS
TRYPSIN
VEGETABLES