Abstract:
:The mammalian bradykinin-degrading enzyme aminopeptidase P (AP-P; E. C. 3.4.11.9) is a metal-dependent enzyme and is a member of the peptidase clan MG. AP-P exists as membrane-bound and cytosolic forms, which represent distinct gene products. A partially truncated clone encoding the cytosolic form was obtained from a human pancreatic cDNA library and the 5' region containing the initiating Met was obtained by 5' rapid accumulation of cDNA ends (RACE). The open reading frame encodes a protein of 623 amino acids with a calculated molecular mass of 69,886 Da. The full-length cDNA with a C-terminal hexahistidine tag was expressed in Escherichia coli and COS-1 cells and migrated on SDS-PAGE with a molecular mass of 71 kDa. The expressed cytosolic AP-P hydrolyzed the X-Pro bond of bradykinin and substance P but did not hydrolyze Gly-Pro-hydroxyPro. Hydrolysis of bradykinin was inhibited by 1,10-phenanthroline and by the specific inhibitor of the membrane-bound form of mammalian AP-P, apstatin. Inductively coupled plasma atomic emission spectroscopy of AP-P expressed in E. coli revealed the presence of 1 mol of manganese/mol of protein and insignificant amounts of cobalt, iron, and zinc. The enzymatic activity of AP-P was promoted in the presence of Mn(II), and this activation was increased further by the addition of glutathione. The only other metal ion to cause slight activation of the enzyme was Co(II), with Ca(II), Cu(II), Mg(II), Ni(II), and Zn(II) all being inhibitory. Removal of the metal ion from the protein was achieved by treatment with 1,10-phenanthroline. The metal-free enzyme was reactivated by the addition of Mn(II) and, partially, by Fe(II). Neither Co(II) nor Zn(II) reactivated the metal-free enzyme. On the basis of these data we propose that human cytosolic AP-P is a single metal ion-dependent enzyme and that manganese is most likely the metal ion used in vivo.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Cottrell GS,Hooper NM,Turner AJdoi
10.1021/bi001585csubject
Has Abstractpub_date
2000-12-12 00:00:00pages
15121-8issue
49eissn
0006-2960issn
1520-4995pii
bi001585cjournal_volume
39pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Fluorescein 5'-isothiocyanate (FITC) was used to modify the lysine residues of bovine transducin (T), a GTP-binding protein involved in phototransduction of rod photoreceptor cells. The incorporation of FITC showed a stoichiometry of approximately 1 mol of FITC/mol of transducin. The labeling was specific for the T al...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00380a022
更新日期:1987-03-24 00:00:00
abstract::Heparanase is a beta-D-endoglucuronidase that cleaves heparan sulfate (HS) and has been implicated in many important physiological and pathological processes, including tumor cell metastasis, angiogenesis, and leukocyte migration. We report herein the identification of active-site residues of human heparanase. Using P...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002080p
更新日期:2000-12-26 00:00:00
abstract::The coexistence effects of multiple kinds of oxidized deoxyribonucleotides were examined using an SV40 origin-dependent in vitro replication system with a HeLa extract. Oxidized dGTP and dATP, 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP) and 2-hydroxy-2'-deoxyadenosine 5'-triphosphate (2-OH-dATP), were used...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062117r
更新日期:2007-06-05 00:00:00
abstract::First identified on the surface of lymphoids as a type II transmembrane protein, CD38 has now been established to have dual functions not only as a receptor but also as a multifunctional enzyme,catalyzing the synthesis of and hydrolysis of a general calcium messenger molecule, cyclic ADP-ribose(cADPR). The receptorial...
journal_title:Biochemistry
pub_type: 杂志文章
doi:
更新日期:2008-12-30 00:00:00
abstract::Hydrogen exchange of the individual amide protons of alanine-90 (F5), glutamine-91 (F6), serine-92 (F7), and histidine-93 (F8) residues in cyanometmyoglobin of sperm whale has been studied by 1H nuclear magnetic resonance spectroscopy at 360 MHz. The amide proton resonance of F5, F6, and F7 have been assigned by use o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00347a024
更新日期:1985-12-17 00:00:00
abstract::Isotope exchange studies show that beef liver fructokinase has a random kinetic mechanism in which release of fructose from the enzyme is slower than that catalytic reaction. The stickiness of fructose in the presence of MgATP is confirmed by isotope partition studies, which show it to be released 0.53 times as fast a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00629a021
更新日期:1977-05-17 00:00:00
abstract::Many bacteria produce isonitrile-containing natural products that are derived from aromatic amino acids. The synthetic clusters that control biosynthesis most commonly encode two enzymes, designated PvcA and PvcB, as well as additional enzymes that direct synthesis of the natural product. The PvcA enzyme installs the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00255
更新日期:2015-04-28 00:00:00
abstract::Cytochrome bc(1) is an integral membrane protein complex essential to cellular respiration and photosynthesis. The Q cycle reaction mechanism of bc(1) postulates a separated quinone reduction (Q(i)) and quinol oxidation (Q(o)) site. In a complete catalytic cycle, a quinone molecule at the Q(i) site receives two electr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0341814
更新日期:2003-08-05 00:00:00
abstract::The nuclear transport of the chromatin remodeling protein nucleoplasmin and chromatin building histones is mediated by importins. Nucleoplasmin (NP) contains a classical bipartite nuclear localization signal (NLS) that is recognized by the importin α/β heterodimer, while histones present multiple NLS-like motifs that ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2008867
更新日期:2011-08-23 00:00:00
abstract::The structures of four isomorphous crystals of ternary complexes of chicken heart citrate synthase with D- or L-malate and acetyl coenzyme A or carboxymethyl coenzyme A have been determined by X-ray crystallography and fully refined at 1.9-A resolution. The structures show that both L-malate and D-malate bind in a ver...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00238a028
更新日期:1991-06-18 00:00:00
abstract::N-(1-Pyrenyl)iodoacetamide has been used to introduce fluorescent probes into Escherichia coli RNA polymerase. After an incubation time of 15 min, approximately 2 pyrene equiv was introduced per enzyme molecule. There was no further increase in modification after more extended periods of incubation. Neither calf thymu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00215a028
更新日期:1991-01-08 00:00:00
abstract::Urea lesions are formed in DNA because of free radical damage of the thymine base, and their occurrence in DNA blocks DNA polymerases, which has deleterious consequences. Recently, it has been shown that urea is capable of forming hydrogen bonding and stacking interactions with nucleobases, which are responsible for t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b00309
更新日期:2016-10-11 00:00:00
abstract::Energy coupling factor (ECF) transporters take up micronutrients in Bacteria and Archaea. They consist of a membrane-embedded S-component that provides substrate specificity and a three-subunit ECF module that couples ATP hydrolysis to transport. The S-components ThiT (for thiamin) and NiaX (for niacin) from Lactococc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00609
更新日期:2015-08-11 00:00:00
abstract::Arachidonic acid metabolism by lipoxygenases and cytochrome P450 monooxygenases produces regioisomeric hydroperoxyeicosatetraenoic acids (HPETEs), hydroxyeicosatetraenoic acids (HETEs), epoxyeicosatrienoic acids (EETs), and dihydroxyeicosatrienoic acids (DHETs), which serve as components of cell signaling cascades. In...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001602y
更新日期:2001-01-30 00:00:00
abstract::Pyrroloquinoline quinone (PQQ) is a small, redox active molecule that serves as a cofactor for several bacterial dehydrogenases, introducing pathways for carbon utilization that confer a growth advantage. Early studies had implicated a ribosomally translated peptide as the substrate for PQQ production. This study pres...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201763d
更新日期:2012-03-20 00:00:00
abstract::The mechanism of binding of reduced 1-(2-carboxyphenylamino)-1-deoxyribulose 5-phosphate (rCdRP) to two different binding sites on the bifunctional enzyme is determined by kinetic studies, using temperature-jump and stopped-flow equipment with fluorescence detection. Two rapid binding processes and a comparatively slo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00593a030
更新日期:1979-12-25 00:00:00
abstract::The interfacial interactions of cholesterol with sphingomyelins (SMs) containing various homogeneous acyl chains have been investigated by Langmuir film balance approaches. Low in-plane elasticity among the packed lipids was identified as an important physical feature of the cholesterol-sphingomyelin liquid-ordered ph...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002791n
更新日期:2001-05-22 00:00:00
abstract::In the cyanobacterium Anabaena, the precursor to tRNA(Leu) has a 249-nucleotide group I intron inserted between the wobble and second bases of the anticodon; the intron self-splices during transcription in vitro [Xu, M. Q., Kathe, S. D., Goodrich-Blair, H., Nierzwicki-Bauer, S. A., & Shub, D. A. (1990) Science 250, 15...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00082a016
更新日期:1993-08-10 00:00:00
abstract::Ricin A-chain (RTA) catalyzes the hydrolytic depurination of a specific adenosine at position 4324 of 28S rRNA. Kinetic isotope effects on the hydrolysis of a small 10mer stem-tetraloop oligonucleotide substrate established the mechanism of the reaction as D(N)*A(N), involving an oxacarbenium ion intermediate in a hig...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0498499
更新日期:2004-05-04 00:00:00
abstract::The role of the 2'-hydroxyl group in RNA--protein interaction has been investigated using MS2 coat protein and its hairpin RNA operator as a model system. Derivatives of the MS2 translational operator were prepared where individual riboses were replaced by deoxyribose and their binding affinities to MS2 coat protein w...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00038a033
更新日期:1995-09-26 00:00:00
abstract::The association-dissociation kinetics of ribosomes from Escherichia coli have been studied under various conditions in a light-scattering stopped-flow apparatus. The dissociation reaction at 2 mg/ml at 25 degrees C, induced by lowering the MgCl2 concentration from 18 to 3 mM, can best be described by three independent...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00671a010
更新日期:1976-12-28 00:00:00
abstract::The three-dimensional structure of a 42-residue fragment containing the N-terminal EGF-like module of blood coagulation factor X was determined by means of 2D NMR spectroscopy and computer simulation. The spectroscopic data consisted of 370 NOE distances and 27 dihedral angle constraints. These were used to generate p...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00141a004
更新日期:1992-07-07 00:00:00
abstract::Processing bodies (P-bodies) are cytoplasmic ribonucleoprotein (RNP) granules primarily composed of translationally repressed mRNAs and proteins related to mRNA decay, suggesting roles in post-transcriptional regulation. P-bodies are conserved in eukaryotic cells and exhibit properties of liquid droplets. However, the...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/acs.biochem.7b01162
更新日期:2018-05-01 00:00:00
abstract::Time-resolved resonance Raman spectroscopy has been employed to monitor geminate heme-CO rebinding in photolyzed HbCO. The excitation frequency was tuned to enhance the scattering from rebound heme sites 20-500 ns subsequent to CO photolysis. The behavior of vFe-C during ligand rebinding has important ramifications co...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00056a017
更新日期:1993-02-09 00:00:00
abstract::A panel of six naïve 14-residue random peptide libraries displayed polyvalently on M13 phage was pooled and sorted against human leukemia inhibitory factor (LIF). After four rounds of selection, a single large family of peptides with the consensus sequence XCXXXXG(A/S)(D/E)(W/F)WXCF was found to bind specifically to L...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035303v
更新日期:2003-11-18 00:00:00
abstract::We have studied the pathway and kinetics of refolding of recombinant human brain-derived neurotrophic factor (BDNF), which is a very tightly-associated dimer in its native state. When BDNF unfolded in 6 M guanidine hydrochloride is diluted 20-fold into phosphate-buffered saline, a partially folded intermediate is rapi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00091a036
更新日期:1993-10-12 00:00:00
abstract::The equilibrium unfolding reaction of Ltn, a metamorphic C-class chemokine, was monitored by tryptophan fluorescence to determine unfolding free energies. Measurements revealed that addition of 150 mM NaCl stabilized the Ltn chemokine fold by approximately 1 kcal/mol. Specific mutations involving Arg23 and Arg43 also ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300842j
更新日期:2012-11-13 00:00:00
abstract::3-Deoxy-d-manno-octulosonate 8-phosphate (KDO8P) synthase catalyzes the condensation of arabinose 5-phosphate (A5P) and phosphoenolpyruvate (PEP) to form KDO8P, a key precursor in the biosynthesis of the endotoxin of Gram-negative bacteria. Earlier studies have established that the condensation occurs with a syn addit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901341h
更新日期:2009-12-15 00:00:00
abstract::A striking feature of the CLIC (chloride intracellular channel) protein family is the ability of its members to convert between a soluble state and an integral membrane channel form. Direct evidence of the structural transition required for the CLIC protein to autonomously insert into the membrane is lacking, largely ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100111c
更新日期:2010-06-29 00:00:00
abstract::Cofilin is an actin regulatory protein that binds to both monomeric and filamentous actin, and has filament severing activity. Although crystal structures for the monomeric forms of both G-actin and cofilin have been described, the structure of the binary cofilin-G-actin complex is not available. Synchrotron protein f...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0121104
更新日期:2002-05-07 00:00:00