Abstract：
:Cytochrome P450BM-3 is a fatty acid monooxygenase that contains the catalytic P450 heme domain covalently attached to a diflavin P450 reductase domain. The function of the linker region connecting the C-terminal end of the heme domain to the N-terminal end of the reductase domain has been studied by deleting parts of the linker and changing the sequence of the linker. Deleting three or six residues or changing an Arg-Lys-Lys stretch in the middle of the linker to Ala-Ala-Ala does not alter the functional properties of either domain. The mutants retain full cytochrome c and ferricyanide reductase activities characteristic of the P450 reductase domain. The heme domain in the mutants retains its ability to bind a fatty acid substrate giving the full low-to-high spin shift and exhibits the normal 450 nm absorption band characteristic of the reduced carbon monoxide complex. However, the six amino acid deletion mutant exhibit nearly undetectable levels of fatty acid hydroxylase activity, the three amino acid deletion mutant about 10% activity, and the three Ala substitution mutant about 50% activity. The mutants also exhibit slower rates of reductase-to-heme electron transfer rates that correlate with the loss in fatty acid hydroxylase activity. These results indicate that the length of the linker and, to a much less extent, the sequence are important for correctly orienting the reductase and heme domains, which apparently is necessary to achieve efficient reductase-to-heme electron transfer rates.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Govindaraj S,Poulos TLdoi
10.1021/bi00035a031subject
Has Abstractpub_date
1995-09-05 00:00:00pages
11221-6issue
35eissn
0006-2960issn
1520-4995journal_volume
34pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:4-Chlorobenzoyl-coenzyme A (4-CBA-CoA) dehalogenase catalyzes the hydrolysis of 4-CBA-CoA to 4-hydroxybenzoyl-coenzyme A (4-HBA-CoA) via a nucleophilic aromatic substitution pathway involving the participation of an active site carboxylate side chain in covalent catalysis. In this paper we report on the identification...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9609533
更新日期：1996-08-20 00:00:00
abstract：:Picosecond transient absorption spectra of rhodopsin and isorhodopsin were measured at room temperature with a double-beam laser spectrophotometer after excitation at 355 nm. Photolysis studies were performed on rhodopsin solubilized in two different detergents (digitonin and 3-[(3-cholamidopropyl)dimethylammonio]-1-p...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00312a008
更新日期：1984-08-14 00:00:00
abstract：:Haloalkane dehalogenase converts haloalkanes to their corresponding alcohols and halides. The reaction mechanism involves the formation of a covalent alkyl-enzyme complex which is hydrolyzed by water. The active site is a hydrophobic cavity buried between the main domain and the cap domain of the enzyme. The enzyme ha...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi952904g
更新日期：1996-05-07 00:00:00
abstract：:Characterization of the complete gene sequence encoding the alpha-galactosidase from Phanerochaete chrysosporium confirms that this enzyme is a member of glycosyl hydrolase family 27 [Henrissat, B., and Bairoch, A. (1996) Biochem. J. 316, 695-696]. This family, together with the family 36 alpha-galactosidases, forms g...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0008074
更新日期：2000-08-15 00:00:00
abstract：:Activated receptors with a tyrosine kinase activity induce a variety of responses like changes in the differentiation and mitogenic status of cells. These responses are mediated in part by p21ras. Some of these activated receptors induce in certain cell types a pronounced, but transient, increase in Ras-GTP. We have s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00189a053
更新日期：1994-06-14 00:00:00
abstract：:A new rapid procedure for purifying 10-formyltetrahydrofolate dehydrogenase results in 90 mg of pure enzyme from two rabbit livers. This abundant liver enzyme is known to bind its product tetrahydropteroylpentaglutamate (H4PteGlu5) so tightly that it does not dissociate during size exclusion chromatography. 10-Formylt...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9619684
更新日期：1996-12-10 00:00:00
abstract：:The reactivity of the mitochondrial (pI = 5) isoenzyme of horse liver aldehyde dehydrogenase was determined by studying the effects of pH on steady-state velocity, burst magnitude, and molecular weight of the enzyme in the absence and presence of Mg2+ ions. The Mg2+ ion activation of the steady-state velocity at pH 7....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00524a049
更新日期：1981-10-13 00:00:00
abstract：:High-resolution proton nuclear magnetic resonance (1H NMR) measurements were made on myosin, heavy meromyosin (HMM), myosin subfragment 1 (S1), light meromyosin (LMM), and actin. A strong signal from amino acid side chains undergoing motions too fast to be accounted for by simple rotations of groups on a rigid backbon...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00586a031
更新日期：1979-09-18 00:00:00
abstract：:Bacillus subtilis CsoR (Bsu CsoR) is a copper-sensing transcriptional repressor that regulates the expression of the copZA operon encoding a copper chaperone and a Cu efflux P-type ATPase, respectively. Bsu CsoR is a homologue of Mycobacterium tuberculosis CsoR (Mtb CsoR), representative of a large Cu(I)-sensing regul...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi900115w
更新日期：2009-04-21 00:00:00
abstract：:The pre-steady-state kinetics of the trichodiene synthase reaction were investigated by rapid chemical quench methods. The single-turnover rate was found to be 3.5-3.8 s-1, a rate 40 times faster than the steady-state catalytic rate (kcat = 0.09 s-1) for trichodiene synthase-catalyzed conversion of farnesyl diphosphat...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi963018o
更新日期：1997-07-08 00:00:00
abstract：:We have used resonance energy transfer to study the spatial relationship between Cys-98 of rabbit skeletal troponin C and Cys-133 of rabbit skeletal troponin I in the reconstituted ternary troponin complex. The donor was introduced by labeling either troponin C or troponin I with N-(iodoacetyl)-N'-(5-sulfo-1-naphthyl)...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00440a029
更新日期：1989-07-11 00:00:00
abstract：:The mechanisms for substrate recognition by two cytoplasmic protein tyrosine phosphatases, PTP-5 and rrbPTP-1, were investigated. Phosphorylation sites on tyrosine-phosphorylated casein, a model PTP substrate, were characterized. Two peptides based on casein phosphorylation sites and one peptide based on the tyrosine ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00097a019
更新日期：1993-11-23 00:00:00
abstract：:Salts and polyamines have a variety of effects on the physical properties of DNA, including stabilization against thermal melting. We wished to gain greater insight into the mechanism of this stabilization by ascertaining its effect on the dynamics of base opening and closing reactions, as measured by NMR. Since the b...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00404a018
更新日期：1988-02-23 00:00:00
abstract：:Transporters from the SLC13 family couple the transport of two to four Na+ ions with a di- or tricarboxylate, such as succinate or citrate. We have previously modeled mammalian members of the SLC13 family, including the Na+/dicarboxylate cotransporter NaDC1 (SLC13A2), based on a structure of the bacterial homologue Vc...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00503
更新日期：2017-08-22 00:00:00
abstract：:Cysteine desulfurases (CDs) are pyridoxal-5'-phosphate (PLP)-dependent enzymes that cleave sulfur from cysteine via an enzyme cysteinyl persulfide intermediate. In vitro studies of these enzymes have generally employed dithiothreitol as a cosubstrate to reductively cleave the persulfide intermediate, and it has been s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi049143e
更新日期：2004-09-28 00:00:00
abstract：:Distinct structural features of DNA, such as the curvature of dA tracts, are important in the recognition, packaging, and regulation of DNA. Physiologically relevant concentrations of magnesium have been found to enhance the curvature of dA tract DNAs, as monitored by solution-state NMR, indicating that the structure ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi010853j
更新日期：2001-08-07 00:00:00
abstract：:The toxin RelE is a ribosome-dependent endoribonuclease implicated in diverse cellular processes, including persistence. During amino acid starvation, RelE inhibits translation by cleaving ribosomal A-site mRNA. Although RelE is structurally similar to other microbial endoribonucleases, the active-site amino acid comp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi401325c
更新日期：2013-12-03 00:00:00
abstract：:Reconstitution of highly purified aquaporin CHIP (channel-forming integral protein) into proteoliposomes was previously shown to confer high osmotic water permeability (Pf) to the membranes [Zeidel et al. (1992) Biochemistry 31, 7436-7440]. Here we report detailed ultrastructural, pharmacologic, and transport studies ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00172a042
更新日期：1994-02-15 00:00:00
abstract：:Coumermycin A(1) is a member of the aminocoumarin family of antibiotics. Unlike its structural relatives, novobiocin and clorobiocin, coumermycin A(1) is a dimer built on a 3-methyl-2,4-dicarboxypyrrole scaffold and bears two decorated noviose sugar components which are the putative target binding motifs for DNA gyras...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi048457z
更新日期：2004-11-30 00:00:00
abstract：:The formation of ternary complexes involving serine proteases, borate, and an alcohol has important implications for understanding the physiological actions of borate and for the development of tight binding inhibitors for this class of enzymes. Recent studies of a related enzyme, gamma-glutamyl transpeptidase, which ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi025583z
更新日期：2002-05-14 00:00:00
abstract：:The photocycle of 124 kDa phytochrome A from Avena sativa was studied by Fourier-transform infrared spectroscopy at low temperatures. Difference spectra between the parent state Pr and the intermediates of the Pr-->Pfr pathway, i.e. lumi-R, meta-Ra, and meta-Rc, and between Pfr and the intermediates of the Pfr-->Pr pa...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960960r
更新日期：1996-08-20 00:00:00
abstract：:Single-molecule fluorescence resonance energy transfer (smFRET) methods were used to study the assembly pathway and DNA unwinding activity of the bacteriophage T4 helicase-primase (primosome) complex. The helicase substrates used were surface-immobilized model DNA replication forks "internally" labeled in the duplex r...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi400231s
更新日期：2013-05-07 00:00:00
abstract：:The DNA binding domain (DBD) of p53 folds by a complex mechanism that involves parallel pathways and multiple intermediates, both on- and off-pathway. This heterogeneity renders DBD particularly susceptible to misfolding and aggregation. The origins of parallel folding mechanisms are not well understood. DBD folding h...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi062106y
更新日期：2007-03-13 00:00:00
abstract：:Deazapurine-containing secondary metabolites comprise a broad range of structurally diverse nucleoside analogues found throughout biology, including various antibiotics produced by species of Streptomyces bacteria and the hypermodified tRNA bases queuosine and archaeosine. Despite early interest in deazapurines as ant...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi900400e
更新日期：2009-05-12 00:00:00
abstract：:Type II dihydrofolate reductases (DHFRs) encoded by the R67 and R388 plasmids are different both in sequence and in structure from known chromosomal DHFRs. These plasmid-derived DHFRs are responsible for conferring trimethoprim resistance to the host strain. A derivative of R388 DHFR, RBG200, has been cloned and overp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00494a011
更新日期：1990-10-23 00:00:00
abstract：:PQQ is an exogenous, tricyclic, quino-cofactor for a number of bacterial dehydrogenases. The final step of PQQ formation is catalyzed by PqqC, a cofactorless oxidase. This study focuses on the activation of molecular oxygen in an enzyme active site without metal or cofactor and has identified a specific oxygen binding...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1015474
更新日期：2011-03-08 00:00:00
abstract：:The bacteriophage T7 DNA helicase/primase (gene 4 protein) is a ring-like hexamer that encircles ssDNA and requires forked DNA to catalyze DNA unwinding. We report that optimal rates of unwinding of forked DNA require ssDNA tails of 55 nucleotides on the 5'-to-3' strand and 15 nucleotides on the 3'-to-5' strand. Surpr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi971644v
更新日期：1997-11-18 00:00:00
abstract：:Deuterium-labeled dihydroorotates bearing one, two, or three deuteriums at the pair of C4 and C5 positions have been synthesized in high isotopic and chiral purity and characterized by NMR and mass spectroscopy. These substrates have been used with the FMN-containing biosynthetic dihydroorotate oxidase from Crithidia ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00307a033
更新日期：1984-06-05 00:00:00
abstract：:The organization of the higher order structure of chromatin has been examined in chicken erythrocyte. Chromatin solubilized during the time course of a gentle micrococcal nuclease digestion of nuclei shows a continuous variation in the distribution of molecular weights. Electron microscopy studies of large chromatin f...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00553a002
更新日期：1980-06-10 00:00:00
abstract：:Significant concentrations (1-10 mM) of nickel(II) were found in solution after incubation of the potent carcinogen nickel subsulfide in 0.05 M Tris-HCl, pH 7.4, solutions containing DNA, rat liver microsomes, and NADPH. The presence of NADPH decreased the rate of solubilization of nickel subsulfide. The solubilized n...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00533a030
更新日期：1982-02-16 00:00:00
