Abstract:
:Recombinant human ferrochelatase has been expressed in Escherichia coli and purified to homogeneity. Metal analyses revealed approximately 2 mol of non-heme Fe per mol of the purified enzyme (M(r) = 40,000). The UV-visible absorption spectrum of the purified enzyme consists of a protein absorption at 278 nm (epsilon approximately 90,000 M-1 cm-1) and bands at 330 nm (epsilon approximately 24,000 M-1 cm-1), 460 nm (shoulder, epsilon approximately 11,000 M-1 cm-1), and 550 nm (shoulder, epsilon approximately 9000 M-1 cm-1) that are indicative of a [2Fe-2S]2+ cluster. The spectra show an additional band at 415 nm that varied in intensity for different preparations and is attributed, at least in part, to a minor component of enzyme-associated high-spin Fe(III) heme. The presence of a single [2Fe-2S]2+,+ cluster as a redox active component of human ferrochelatase was confirmed by variable-temperature MCD and EPR studies of the dithionite-reduced enzyme which showed the presence of a S = 1/2 [2Fe-2S]+ cluster in addition to residual high spin Fe(II) heme. The reduced enzyme exhibits a S = 1/2 EPR signal, g = 2.00, 1.94, 1.91 accounting for 0.75 +/- 0.25 spins/molecule, that readily saturates at low microwave powers below 10 K but is observable without significant broadening at temperatures up to 100 K. The Fe-S cluster is labile and gradually disappears over period of 24 h, with concomitant loss of enzyme activity, when the enzyme is stored aerobically at 4 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Dailey HA,Finnegan MG,Johnson MKdoi
10.1021/bi00168a003subject
Has Abstractpub_date
1994-01-18 00:00:00pages
403-7issue
2eissn
0006-2960issn
1520-4995journal_volume
33pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The thermodynamic quantities of the anti-dinitrophenyl antibody-hapten interaction are reported for rabbit, goat, and guinea pig antibodies. Rabbit and goat antibodies had similar exothermic enthalpy changes for their reaction with 2,4-dinitrophenyl-L-lysine (-13.9 and -14.8 kcal/mol, respectively). The enthalpy chang...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00694a032
更新日期:1975-11-18 00:00:00
abstract::The first transmembrane (TM1) helix in the red cell anion exchanger (AE1, Band 3, or SLC4A1) acts as an internal signal anchor that binds the signal recognition particle and directs the nascent polypeptide chain to the endoplasmic reticulum (ER) membrane where it moves from the translocon laterally into the lipid bila...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b00966
更新日期:2017-02-07 00:00:00
abstract::The biodegradative 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase of Pseudomonas mevalonii catalyzes the NAD(+)-dependent conversion of (S)-HMG-CoA to (R)-mevalonate. Crystallographic analysis of abortive ternary complexes of this enzyme revealed lysine 267 located at a position in the active site, suggesti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9902687
更新日期:1999-07-13 00:00:00
abstract::Proton NMR studies were conducted on the complementary 15-mer duplex d(5'-TACTCTTCTTGACCT).(5'-AGGTCAAGAAGAGTA) (designated as unmodified 15-mer duplex) spanning a portion of the mouse c-Ha-ras protooncogene centered around codon 61. Identical studies were carried out on the same sequence, after specific modification ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00155a011
更新日期:1992-10-13 00:00:00
abstract::The light-induced QA-/QA FTIR difference spectrum of the photoreduction of the primary quinone (QA) in reaction centers (RCs) from Rhodobacter sphaeroides exhibits a set of complex differential bands between 1750 and 1715 cm(-1). Several of these features correspond in frequency to bands that bleach in the HA-/HA FTIR...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962871k
更新日期:1997-04-15 00:00:00
abstract::Complete assignments of nitrogen-15 resonances of gramicidin S have been made in dimethyl sulfoxide, trifluoroethanol, and in a solvent mixture of dimethyl sulfoxide (50%) and methanol (50%). The assignments are achieved by utilizing the secondary structure of gramicidin S, by comparing the nitrogen-15 spectrum of gra...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00606a005
更新日期:1978-06-27 00:00:00
abstract::Three unfolding domains in rabbit muscle aldolase destabilized in 3 M urea have been identified from their unfolding rate constants (0.10, 0.036, and 0.0064 min-1). The populations of folded and various, partially unfolded forms were determined by amide hydrogen exchange and mass spectrometry. Results of this study sh...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972711o
更新日期:1998-05-05 00:00:00
abstract::The Sir2 enzyme family is responsible for a newly classified chemical reaction, NAD(+)-dependent protein deacetylation. New peptide substrates, the reaction mechanism, and the products of the acetyl transfer to NAD(+) are described for SIR2. The final products of SIR2 reactions are the deacetylated peptide and the 2' ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011858j
更新日期:2001-12-25 00:00:00
abstract::Zinc, the catalytically essential metal of angiotensin converting enzyme (ACE), has been replaced by cobalt(II) to give an active, chromophoric enzyme that is spectroscopically responsive to inhibitor binding. Visible absorption spectroscopy and magnetic circular dichroic spectropolarimetry have been used to character...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00397a014
更新日期:1987-11-17 00:00:00
abstract::Respiratory complex I couples the electron transfer exclusively from NADH to a quinone with the translocation of protons across the membrane. However, Escherichia coli adapts to imposed high cellular NADPH concentrations by selecting the mutations E183A(F) and E183G(F) that lead to a high catalytic efficiency of compl...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00160
更新日期:2015-05-12 00:00:00
abstract::Detection of specific reaction products is a powerful approach for dissecting out pathways that mediate oxidative damage in vivo. Eosinophil peroxidase (EPO), an abundant protein secreted from activated eosinophils, has been implicated in promoting oxidative tissue injury in conditions such as asthma, allergic inflamm...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982401l
更新日期:1999-03-23 00:00:00
abstract::Amphiphiles respond both to polar and to nonpolar solvents. In this paper X-ray diffraction and osmotic stress have been used to examine the phase behavior, the structural dimensions, and the work of deforming the monolayer-lined aqueous cavities formed by mixtures of dioleoylphosphatidylethanolamine (DOPE) and dioleo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00453a010
更新日期:1990-01-09 00:00:00
abstract::Protein splicing is a self-catalyzed post-translational modification in which the intein enzyme excises itself from a precursor protein and ligates the flanking sequences to produce a mature protein. We report the solution structure of a 136-residue DnaX mini-intein enzyme derived from the cyanobacterium Spirulina pla...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00828
更新日期:2020-12-22 00:00:00
abstract::Four new in vivo metabolites of vitamin D3 were isolated from the blood plasma of chicks given large doses of vitamin D3. The metabolites were isolated by methanol-chloroform extraction and a series of chromatographic procedures. By use of mass spectrometry, ultraviolet absorption spectrophotometry, and specific chemi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00529a010
更新日期:1981-12-22 00:00:00
abstract::The O-linked protein glycosylation pathway in Neisseria gonorrhoeae is responsible for the synthesis of a complex oligosaccharide on undecaprenyl diphosphate and subsequent en bloc transfer of the glycan to serine residues of select periplasmic proteins. Protein glycosylation (pgl) genes have been annotated on the bas...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2003372
更新日期:2011-06-07 00:00:00
abstract::The ovalbumin (Ov) gene contains a number of regulatory elements that control its transcriptional activity and restrict expression to avian oviduct. One major regulatory region, the steroid-dependent regulatory element (SDRE), is required for induction by estrogen and corticosterone. Another region, the negative regul...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0005862
更新日期:2000-07-25 00:00:00
abstract::Porcine aortic myosin is a smooth muscle contractile protein similar to its striated muscle counterpart. Electrophoresis in sodium dodecyl sulfate indicates that the molecule consists of three classes of subunits with polypeptide chain molecular weights of 192,000, 19,000, and 15,000. At 277 nm the absorption spectrum...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00576a003
更新日期:1979-05-01 00:00:00
abstract::beta-amyloid peptide (Abeta) is the primary constituent of senile plaques, a defining feature of Alzheimer's disease. Aggregated Abeta is toxic to neurons, but the mechanism of toxicity remains unproven. One proposal is that Abeta toxicity results from relatively nonspecific Abeta-membrane interactions. We hypothesize...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0001980
更新日期:2000-08-22 00:00:00
abstract::An issue that is central to understanding cellular fatty acid (FA) metabolism is whether physiologic transport of FA across cell membranes requires protein mediation or can be satisfied by the rate of spontaneous movement through the lipid phase. For this reason, considerable effort has been devoted to determining the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962007s
更新日期:1997-05-13 00:00:00
abstract::On the basis of the available high-resolution structures of mandelate racemase (MR) from Pseudomonas putida [Landro, J. A., Gerlt, J. A., Kozarich, J. W., Koo, C. W., Shah, V. J., Kenyon, G. L., Neidhart, D. J., Fujita, J., & Petsko, G. A. (1994) Biochemistry 33, 635-643], Lys 166 and His 297 are positioned appropriat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00009a007
更新日期:1995-03-07 00:00:00
abstract::The regulation of the rate-controlling enzyme in cholesterol biosynthesis and of the incorporation of [14C]oleate into cholesterol esters were studied in established lymphoid cell lines from normal subjects and compared with that of eight patients with genetic abnormalities of lipid metabolism. The activity of 3-hydro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00648a011
更新日期:1976-02-10 00:00:00
abstract::The function of Lys-59, Arg-141, and Arg-277 in PAPS binding and catalysis of the flavonol 3-sulfotransferase was investigated. Affinity chromatography of conservative mutants with PAPS analogues allowed us to determine that Lys-59 interacts with the 5' portion of the nucleotide, while Arg-141 interacts with the 3' po...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982239m
更新日期:1999-03-30 00:00:00
abstract::Pantothenate synthetase (PS, EC 6.3.2.1) is the last enzyme in the pantothenate biosynthesis pathway, a metabolic pathway identified as a potential target for new antimicrobials. PS catalyzes the ATP-dependent condensation of pantoate and beta-alanine to form pantothenate. Here we report the overexpression, purificati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1004206
更新日期:2010-08-03 00:00:00
abstract::The l-lyxonate dehydratase (LyxD) in vitro enzymatic activity and in vivo metabolic function were assigned to members of an isofunctional family within the mandelate racemase (MR) subgroup of the enolase superfamily. This study combined in vitro and in vivo data to confirm that the dehydration of l-lyxonate is the bio...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi5004298
更新日期:2014-05-27 00:00:00
abstract::The redox properties of the [2Fe-2S] cluster in the 24 kDa subunit of bovine heart mitochondrial NADH:ubiquinone oxidoreductase (complex I) and three of its homologues have been defined using protein-film voltammetry. The clusters in all four examples display characteristic, pH-dependent redox transitions, which, unus...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026026f
更新日期:2002-08-06 00:00:00
abstract::We have studied the tridecadeoxynucleotide CGCGAATTACGCG (I), which contains an additional A at position 9 compared to the dodecanucleotide of which the crystal structure has been determined. Sequence I exhibits no distinct melting curve and also has a concentration-dependent pattern of peaks on reverse-phase chromato...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00371a025
更新日期:1986-11-18 00:00:00
abstract::myo-Inositol oxygenase (MIOX) uses iron as its cofactor and dioxygen as its cosubstrate to effect the unique, ring-cleaving, four-electron oxidation of its cyclohexan-(1,2,3,4,5,6-hexa)-ol substrate to d-glucuronate. The nature of the iron cofactor and its interaction with the substrate, myo-inositol (MI), have been p...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0519607
更新日期:2006-05-02 00:00:00
abstract::During fertilization, free-swimming mouse sperm bind to mZP3 (approximately 83 000 Mr), one of three zona pellucida glycoproteins, and once bound undergo the acrosome reaction, a type of cellular exocytosis [Wassarman, P. M., & Litscher, E. S. (1995) Curr. Top. Dev. Biol. 30, 1-19]. Sperm recognize and bind to specifi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952722m
更新日期:1996-04-02 00:00:00
abstract::The three-dimensional high-resolution structure of rhodopsin is unknown, as is the case for almost all integral membrane patients. As part of an alternative approach to determine of membrane protein structure, we are pursuing the structure of cytoplasmic domains of this G-protein receptor. A peptide, rhoIII, with the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00045a002
更新日期:1995-11-14 00:00:00
abstract::The proline catabolic enzyme Δ(1)-pyrroline-5-carboxylate dehydrogenase (ALDH4A1) catalyzes the NAD(+)-dependent oxidation of γ-glutamate semialdehyde to l-glutamate. In Saccharomyces cerevisiae, ALDH4A1 is encoded by the PUT2 gene and known as Put2p. Here we report the steady-state kinetic parameters of the purified ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi500048b
更新日期:2014-03-04 00:00:00