Partial reactions and chemical rescue of site-directed mutants of Rubisco as mechanistic probes

Conference · Tue Jan 01 00:00:00 EST 1991

OSTI ID:5756989

Harpel, M R; Larimer, F W; Lee, E H; Mural, R J; Smith, H B; Soper, T S; Hartman, F C

Given the current state of knowledge of the reaction pathways catalyzed by D-ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the elucidation of the three-dimensional structure of several different forms of the enzyme, sit-directed mutagenesis offers the potential to decipher catalytic roles of active-site residues and to unravel the functional significance of various structural elements. Especially intriguing are intersubunit, electrostatic interactions at the active site between Glu48 and Lys168 of the nonactivated (noncarbamylated) enzyme and between Glu48 and Lys329 of the activated (carbamylated) enzyme. In this paper, we describe two approaches to address the roles of electrostatic interactions at the active site and the roles of the participant residues: (1) characterization of pertinent site-directed mutants, including their abilities to catalyze partial reactions and (2) subtle alteration of the active-site microenvironment by manipulation of these proteins with exogenous reagents.

• Research Organization: Oak Ridge National Lab., TN (United States)
• Sponsoring Organization: USDOE; USDOE, Washington, DC (United States)
• DOE Contract Number: AC05-84OR21400
• OSTI ID: 5756989
• Report Number(s): CONF-911295-1; ON: DE92007255
• Resource Relation: Conference: Royal Swedish Academy of Science (RSAS) nobel symposium, Stockholm (Sweden), 4-6 Dec 1991
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
CATALYSTS
PROTEIN ENGINEERING
PROTEINS
CONFIGURATION INTERACTION
CARBOXYLATION
CATALYSIS
ENZYME ACTIVITY
GLUTAMINE
LYSINE
MUTAGENESIS
OXYGENASES
PROTEIN STRUCTURE
REACTION INTERMEDIATES
AMIDES
AMINO ACIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
ENZYMES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXIDOREDUCTASES
400201* - Chemical & Physicochemical Properties
550200 - Biochemistry