Purification and properties of Renilla reniformis luciferase

Journal Article · Tue Jan 11 04:00:00 EST 1977 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00620a014· OSTI ID:7332162

Matthews, J C; Hori, K; Cormier, M J

Luciferase from the anthozoan coelenterate Renilla reniformis (Renilla luciferin:oxygen 2-oxidoreductase (decarboxylating), EC 1.13.12.5.) catalyzes the bioluminescent oxidation of Renilla luciferin producing light (lambda/sub B/ 480 nm, Q/sub B/ 5.5 percent), oxyluciferin, and CO/sub 2/. Using a combination of ion-exchange, molecular-sieve, sulfhydryl-exchange, and affinity chromatography, luciferase has been purified, approximately 12,000-fold with 24 percent recovery, to homogeneity as judged by analysis with disc and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, gel filtration, and ultracentrifugation. Renilla luciferase is active as a nearly spherical single polypeptide chain monomer of 3.5 x 10/sup 4/ daltons having a specific activity of 1.8 x 10/sup 15/ h..nu..s/sup -1/ mg/sup -1/ and a turnover number of 111 ..mu..mol min/sup -/1 ..mu..mol/sup -1/ of enzyme. This enzyme has a high content of aromatic and hydrophobic amino acids such that it has an epsilon/sub 280 nm/ 0.1 percent/ of 2.1 and an average hydrophobicity of 1200 cal residue/sup -1/. The high average hydrophobicity of luciferase, which places it among the more hydrophobic proteins reported, is believed to account, at least in part, for its tendency to self-associate forming inactive dimers and higher molecular weight species.

• Research Organization: Univ. of Georgia, Athens
• OSTI ID: 7332162
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 16:1; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
CENTRIFUGATION
CHEMICAL PROPERTIES
CHROMATOGRAPHY
CNIDARIA
ELECTROPHORESIS
ENZYMES
INVERTEBRATES
LUCIFERASE
LUMINESCENCE
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
OXIDASES
OXIDOREDUCTASES
PROTEINS
PURIFICATION
SEPARATION PROCESSES
ULTRACENTRIFUGATION