Abstract:
:MAP kinase kinase (MAPKK) is a dual specificity protein kinase that phosphorylates and activates MAP kinase in vivo. In this study, four mouse monoclonal single-chain Fv (scFv) antibodies (Y1-6, Y1-7, Y3-6, and Y3-11) that can specifically bind to Xenopus MAPKK were isolated from combinatorial scFv-displaying phage libraries. Three scFv clones (Y1-6, Y1-7, and Y3-6) were shown to efficiently inhibit MAPKK activity in vitro. Point mutation (D98K) at VH-CDR3 of one (Y1-6) of these three clones markedly reduced its neutralizing activity. The wild-type scFv (Y1-6) inhibited the Mos-induced MAP kinase activation and germinal vesicle breakdown when injected into immature Xenopus oocytes, whereas the mutant scFv, Y1-6 (D98K), did not. The three neutralizing scFv clones (Y1-6, Y1-7, and Y3-6) were shown to bind to NH2-terminal residues 1-23 of Xenopus MAPKK, whereas the epitope of a Y3-11 clone with no neutralizing activity was shown to lie between residues 33 and 67 of MAPKK. Furthermore, a synthetic peptide (the N16 peptide) corresponding to residues 2-17 of MAPKK suppressed the neutralizing activity of the wild-type Y1-6, and a rabbit polyclonal antibody against the N16 peptide was found to possess a strong neutralizing activity against MAPKK. These results demonstrate that the neutralizing antibodies characterized here inhibit the kinase activity of MAPKK by binding to the NH2-terminal segment of MAPKK.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Kosako H,Akamatsu Y,Tsurushita N,Lee KK,Gotoh Y,Nishida Edoi
10.1021/bi960956fsubject
Has Abstractpub_date
1996-10-08 00:00:00pages
13212-21issue
40eissn
0006-2960issn
1520-4995pii
bi960956fjournal_volume
35pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Previous studies of P-glycoprotein have demonstrated that its function can be modulated by phosphorylation. In the present study, inhibition of protein kinase C with calphostin C or stauroporine or prolonged treatment with the phorbol ester TPA decreased phosphorylation of P-glycoprotein, and impaired transport of vin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00086a022
更新日期:1993-09-07 00:00:00
abstract::F(1)-ATPase mutations in Escherichia coli that changed the strength of hydrogen bonds between the alpha and beta subunits in a location that links the catalytic site to the interface between the beta catch loop and the gamma subunit were examined. Loss of the ability to form the hydrogen bonds involving alphaS337, bet...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052592+
更新日期:2006-09-19 00:00:00
abstract::CNAD (5-beta-D-ribofuranosylnicotinamide adenine dinucleotide) is an isosteric C-glycosidic analogue of NAD(H) containing a neutral pyridine ring. CPAD (5-beta-D-ribofuranosylpicolinamide adenine dinucleotide) is a closely related pyridine-containing analogue with the pyridine nitrogen on the opposite side of the ring...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00205a009
更新日期:1994-10-04 00:00:00
abstract::Isopentenyl phosphate kinase (IPK) catalyzes the phosphorylation of isopentenyl phosphate to form the isoprenoid precursor isopentenyl diphosphate in the archaeal mevalonate pathway. This enzyme is highly homologous to fosfomycin kinase (FomA), an antibiotic resistance enzyme found in a few strains of Streptomyces and...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201662k
更新日期:2012-01-31 00:00:00
abstract::We have previously shown functional differences in fibrinogen from hyperhomocysteinemic rabbits compared to that in control rabbits. This acquired dysfibrinogenemia is characterized by fibrin clots that are composed of abnormally thin, tightly packed fibers with increased resistance to fibrinolysis. Homocysteine thiol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052076j
更新日期:2006-02-28 00:00:00
abstract::By using some mutant peptides of transcription factor Spl, phosphate backbone contacts with the DNA binding protein containing three zinc fingers have been investigated by alkylation interference, circular permutation, DNase I footprinting, and methylation protection methods. The ethylation interference analyses of Sp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952530r
更新日期:1996-07-02 00:00:00
abstract::The importance of DNA polymerase-DNA minor groove interactions on translesion synthesis (TLS) was examined in vitro using variants of exonuclease-deficient Klenow polymerase and site-specifically modified DNA oligonucleotides. Polymerase variant R668A lacks primer strand interactions, while variant Q849A lacks templat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047591g
更新日期:2005-05-10 00:00:00
abstract::Calpains are intracellular calcium-activated cysteine proteases whose unregulated proteolysis following the loss of calcium homeostasis can lead to acute degeneration during ischemic episodes and trauma, as well as Alzheimer's disease and cataract formation. The determination of the crystal structure of the proteolyti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060425j
更新日期:2006-06-20 00:00:00
abstract::His51 in horse liver alcohol dehydrogenase (ADHE) has been proposed to act as a proton donor/acceptor in the NAD+/NADH-dependent oxidation/reduction of alcohol/aldehyde. The residue corresponding to His51 of ADHE is Val51 (Val45 in the protein sequence) in benzyl alcohol dehydrogenase (BADH) encoded by TOL plasmid pWW...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970726g
更新日期:1998-03-10 00:00:00
abstract::A conformational analysis of the valine side chains of ribonuclease T1 (RNase T1) was performed using NMR spectroscopy, in particular homonuclear (1H, 1H and 13C, 13C) and heteronuclear (1H, 15N and 1H, 13C) vicinal spin-spin coupling constants as obtained from E.COSY-type NMR experiments. The coupling constants relat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00184a017
更新日期:1994-05-10 00:00:00
abstract::Treatment of ADP-glucose pyrophosphorylase (EC 2.7.7.27) from the cyanobacterium Anabaena PCC 7120 with phenylglyoxal in 50 mM Hepes, pH 8.0, at 25 degrees C resulted in a time- and concentration-dependent loss of enzyme activity. Phosphate, the inhibitor, protected the enzyme from inactivation most effectively, while...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9713355
更新日期:1997-10-21 00:00:00
abstract::The crystal structure of the ternary complex citrate synthase-oxaloacetate-carboxymethyl coenzyme A has been solved to a resolution of 1.9 A and refined to a conventional crystallographic R factor of 0.185. The structure resembles a proposed transition state of the condensation reaction and suggests that the condensat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:
更新日期:1990-03-06 00:00:00
abstract::Kinetics of reduction of phototrophic bacterial flavocytochromes c by exogenous flavin semiquinones and fully reduced flavins generated by laser flash photolysis have been studied. The mechanisms of reduction of Chromatium and Chlorobium flavocytochromes c are more similar to one another than previously thought. Neith...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00327a002
更新日期:1985-03-12 00:00:00
abstract::The septins are a conserved family of guanosine-5'-triphosphate (GTP)-binding proteins. In mammals they are involved in a variety of cellular processes, such as cytokinesis, exocytosis, and vesicle trafficking. Specifically, SEPT4 has also been shown to be expressed in both human colorectal cancer and malignant melano...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061549z
更新日期:2006-11-21 00:00:00
abstract::Substitution of cysteine for proline-39 in Escherichia coli dihydrofolate reductase by oligonucleotide-directed mutagenesis positions the new cysteine adjacent to already existing cysteine-85. When the mutant protein is expressed in the E. coli cytosol, the cysteine sulfur atoms are found, by X-ray crystallographic an...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00382a017
更新日期:1987-04-21 00:00:00
abstract::The major covalently linked multimolecular D fragments found in plasmic digests of factor XIIIa cross-linked fibrin formed under physiological pH and ionic strength conditions consist of D dimers, D trimers, and D tetramers. These fragments are linked by epsilon-amino-gamma-glutamyllysine bonds in the carboxy-terminal...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00118a040
更新日期:1992-01-28 00:00:00
abstract::In eukaryotes, a major route for ATP-dependent protein breakdown proceeds through covalent intermediates of target proteins destined for degradation and the highly conserved, 76 amino acid protein ubiquitin. In rabbit reticulocytes, it has been shown that hemin effectively inhibits this pathway by blocking the catabol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00409a025
更新日期:1988-05-03 00:00:00
abstract::In this study, we have investigated the influence of regions outside the DNA-binding domain of the human glucocorticoid receptor on high-affinity DNA binding. We find that the DNA-binding domain shows a 10-fold lower affinity for a palindromic DNA-binding site than the intact receptor. The N-terminal part of the recep...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00152a047
更新日期:1992-09-22 00:00:00
abstract::The Raman laser temperature-jump technique has been used to measure the kinetics of the coil to helix reaction of poly(cytidylic acid) [poly(C)] in aqueous cosolvent mixtures. The rate of helix formation has a low activation energy and is proportional to reciprocal solvent viscosity. The observations suggest helix for...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00509a003
更新日期:1981-03-17 00:00:00
abstract::Escherichia coli DNA photolyase contains 1,5-dihydro-FAD (FADH2) plus 5,10-methenyl-tetrahydrofolate (5,10-CH+-H4folate). Both chromophores are fluorescent, and either can function as a sensitizer in catalysis. At 77 K separate fluorescence emission bands are observed for FADH2 (lambda max = 505 nm, shoulder at 540 nm...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00425a007
更新日期:1988-12-13 00:00:00
abstract::Site-directed spin labeling (SDSL), the site-specific incorporation of nitroxide spin-labels into a protein, has allowed us to investigate ligand-induced conformational changes in the ligand-binding domain of human estrogen receptor alpha (hERalpha-LBD). EPR (electron paramagnetic resonance) spectroscopy of the nitrox...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035566p
更新日期:2004-02-24 00:00:00
abstract::Replication protein A (RPA) is a heterotrimeric protein that is required for DNA replication and most DNA repair pathways. RPA has previously been shown to play a role in recognizing and binding damaged DNA during nucleotide excision repair (NER). RPA has also been suggested to play a role in psoralen DNA interstrand ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi800460d
更新日期:2008-09-23 00:00:00
abstract::Phospholipid transfer protein (PLTP) is expressed by macrophage-derived foam cells in human atherosclerotic lesions, suggesting a regulatory role for PLTP in cellular cholesterol homeostasis. However, the exact role of PLTP in the reverse cholesterol transport pathway is not known. PLTP is present in plasma as two for...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700833h
更新日期:2007-10-23 00:00:00
abstract::Chemosensory cilia isolated from the olfactory epithelium of Rana catesbeiana were solubilized with Lubrol PX in the presence of supplementary lipid, forskolin, and sodium fluoride. Subsequent removal of the detergent by adsorption onto Biobeads SM2 results in the formation of proteoliposomes that display forskolin- a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00417a040
更新日期:1988-08-23 00:00:00
abstract::The rate of 1-palmitoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino] dodecanoyl] phosphatidylcholine (P-C12-NBD-PC) transfer between dioleoylphosphatidylcholine vesicles was measured by a technique based on resonance energy transfer between P-C12-NBD-PC and N-(lissamine rhodamine B sulfonyl)dioleoylphosphatidyletha...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00364a021
更新日期:1986-08-12 00:00:00
abstract::The PLL(PTE-like lactonase)-group of enzymes within the amidohydrolase superfamily hydrolyze N-acyl-homoserine lactones (AHLs) that are involved in bacterial quorum-sensing pathways. These enzymes possess the (beta/alpha)(8)-barrel fold and serve as attractive templates for in vitro evolution and engineering of quorum...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9004045
更新日期:2009-05-26 00:00:00
abstract::AMSH, a deubiquitinating enzyme (DUB) with exquisite specificity for Lys63-linked polyubiquitin chains, is an endosome-associated DUB that regulates sorting of activated cell-surface signaling receptors to the lysosome, a process mediated by the members of the endosomal sorting complexes required for transport (ESCRT)...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi401106b
更新日期:2013-11-05 00:00:00
abstract::Hemophores from Serratia marcescens (HasA(sm)) and Pseudomonas aeruginosa (HasA(p)) bind hemin between two loops, which harbor the axial ligands H32 and Y75. Hemin binding to the Y75 loop triggers closing of the H32 loop and enables binding of H32. Because Yersinia pestis HasA (HasA(yp)) presents a Gln at position 32,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400280z
更新日期:2013-04-23 00:00:00
abstract::The transcriptional activity of the beta-globin genes is regulated by a complex genetic element, the locus control region (LCR), at the 5'-end of the beta-globin locus. Tandem binding sites for the erythroid-specific transcription factor NF-E2 are important for the transcriptional activation function of the LCR. We di...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00050a015
更新日期:1995-12-19 00:00:00
abstract::TAR DNA-binding protein 43 (TDP-43) has been identified as the major constituent of the proteinaceous inclusions that are characteristic of most forms of amyotrophic lateral sclerosis (ALS) and ubiquitin positive frontotemporal lobar degeneration (FTLD). Wild type TDP-43 inclusions are a pathological hallmark of >95% ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.9b00592
更新日期:2019-10-01 00:00:00