Identification of essential lysyl and cysteinyl residues in spinach ribulosebisphosphate carboxylase/oxygenase modified by the affinity label n-bromoacetylethanolamine phosphate

Journal Article · Fri Aug 25 04:00:00 EDT 1978 · J. Biol. Chem.; (United States)

OSTI ID:6624008

Schloss, J V; Stringer, C D; Hartman, F C

We reported earlier that N-bromoacetylethanolamine phosphate is an affinity label for spinach ribulosebisphosphate carboxylase/oxygenase. We now show inactivation to be correlated directly with the alkylation either of a single lysyl residue (in the presence of Mg/sup 2 +/) or of 2 different cysteinyl residues (in the absence of Mg/sup 2 +/), consistent with the likelihood that these residues are located in the active site region. This proposition is further supported by the demonstration that the residues are protected from alkylation by substrate, a competitive inhibitor, or the transition state analog 2-carboxyribitol bisphosphate. Tryptic peptides that contain the modified residues have been isolated and sequenced. One of the 2 cysteinyl residues that are subject to alkylation is only 3 residues distant in sequence from the lysyl residue modified by bromoacetylethanolamine phosphate. This lysyl residue is identical with 1 of the 2 lysyl residues alkylated by the previously described affinity label, 3-bromo-1,4-dihydroxy-2-butanone 1,4-bisphosphate.

• Research Organization: Univ. of Tennessee, Oak Ridge
• OSTI ID: 6624008
• Journal Information: J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 253:16; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
CARBOXYLASE
CARBOXYLIC ACIDS
CHEMICAL ANALYSIS
CYSTEINE
ENZYMES
FOOD
INACTIVATION
KINETICS
LYASES
LYSINE
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXIDOREDUCTASES
OXYGEN COMPOUNDS
PHOSPHATES
PHOSPHORUS COMPOUNDS
REACTION KINETICS
RESIDUES
SPINACH
THIOLS
VEGETABLES