Biomaterial Database

Purification and characterization of cytochrome b5 reductase from the house fly, Musca domestica. 1996

M Zhang, and J G Scott

Department of Entomology, Cornell University, Ithaca, NY 14853-0999, USA.

NADH-cytochrome b5 reductase (b5R) from the house fly was purified through solubilization of microsomes with Triton X-100 followed by DEAE, carboxylmethyl and 5'-ADP affinity column chromatography. Yields of 9% with a 320-fold increase in NADH-ferricyanide reductase specific activity and 2% with a 76-fold increase in NADH-cytochrome b5R specific activity were obtained. Two forms of b5R, a major form with the apparent molecular mass of 31 kDa and a minor form of 33 kDa, were obtained. Both forms of purified b5R could reduce cytochrome b5 and both could use NADH or NADPH as an electron donor, although NADH was more efficient. Kinetics of the b5R activities were also studied. The 31-kDa b5R consists of integral of 291 amino acids with the NH2-terminal sequence of Thr-Ala-Arg-Leu-Arg-Thr-Leu-Ile-Asp-Ala. An antiserum developed against the 31-kDa b5R recognized both forms of b5R. Using this polyclonal antiserum as a probe, immunologically reactive proteins were found in microsomes from five species of Diptera, mouse and rat liver but not in spider mites nor insects from other orders.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D008970	Molecular Weight	The sum of the weight of all the atoms in a molecule.	Molecular Weights,Weight, Molecular,Weights, Molecular
D010084	Oxidation-Reduction	A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).	Redox,Oxidation Reduction
D003579	Cytochrome Reductases		Reductases, Cytochrome
D005292	Ferricyanides	Inorganic salts of the hypothetical acid, H3Fe(CN)6.
D006793	Houseflies	Flies of the species Musca domestica (family MUSCIDAE), which infest human habitations throughout the world and often act as carriers of pathogenic organisms.	Musca domestica,Housefly,Musca domesticas,domesticas, Musca
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000596	Amino Acids	Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.	Amino Acid,Acid, Amino,Acids, Amino
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia