Abstract：
:LTbetaR is a member of the TNF receptor family of proteins. It binds to two different cell surface ligands, LIGHT, a homotypic trimer, and LTalpha1beta2, a heterotypic trimer. We have measured the affinities of the dimeric IgG fusion protein, LTbetaRIgG, and monomeric LTbetaR protein binding to both LIGHT and LTalpha1beta2 using surface plasmon resonance and found values of <0.1 and 38 nM for LIGHT and <0.1 and 48 nM for LTalpha1beta2, respectively. We also determined the stoichiometries of binding for both forms of the receptor LTbetaRIgG and LTbetaR binding to LIGHT. The data obtained from several biophysical methods are consistent with receptor polypeptide to trimeric ligand ratios of 2:1. The determined masses of the complexes using SEC-LS corresponded to a single LTbetaRIgG bound to a LIGHT trimer, or two LTbetaR bound per LIGHT. Sedimentation velocity of varied ratios of LTbetaR to a fixed concentration of LIGHT were analyzed by SEDANAL and were successfully fit with a model with two tight binding sites on LIGHT and one poor affinity site. Isothermal calorimetric titration of LIGHT with either LTbetaR or LTbetaRIgG also demonstrated stoichiometries of 1:2 and 1:1, respectively. The binding of LTbetaR to LIGHT was endothermic and, hence, entropy-driven. TNFR p55 (extracellular domain) complexed with the trimeric ligand, TNFbeta, exhibits a 3:1 receptor/ligand stoichiometry. This complex has been used as the prototypical model setting the receptor-ligand complexation paradigm for the entire TNF family. The LTbetaR/LIGHT binding stoichiometry of 2:1 demonstrated here does not fit the paradigm. This has numerous implications for cell biology including signaling requiring only dimerization of LTbetaR rather than trimerization as expected from the structural paradigm.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Eldredge J,Berkowitz S,Corin AF,Day ES,Hayes D,Meier W,Strauch K,Zafari M,Tadi M,Farrington GKdoi
10.1021/bi060210+subject
Has Abstractpub_date
2006-08-22 00:00:00pages
10117-28issue
33eissn
0006-2960issn
1520-4995journal_volume
45pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Halogen bonding is a weak chemical force that has so far mostly found applications in crystal engineering. Despite its potential for use in drug discovery, as a new molecular tool in the direction of molecular recognition events, it has rarely been assessed in biopolymers. Motivated by this fact, we have developed a p...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00429
更新日期：2017-06-27 00:00:00
abstract：:PII proteins are sensors of alpha-ketoglutarate and adenylylate energy charge that regulate signal transduction proteins, metabolic enzymes, and permeases involved in nitrogen assimilation. Here, purified Escherichia coli PII and two of its receptors, ATase and NRII, were used to study the mechanisms of sensation by P...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9011594
更新日期：2009-12-08 00:00:00
abstract：:The beta-glucosidase from Agrobacterium faecalis (previously Alcaligenes faecalis) has been subjected to a detailed kinetic investigation with a range of substrates to probe its specificity and mechanism. It has a relatively broad specificity for the substrate sugar moiety and exhibits a classical pH dependence for it...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00156a015
更新日期：1992-10-20 00:00:00
abstract：:Factor X (FX) is one of the major players in the blood coagulation cascade. Upon activation to FXa, it converts prothrombin to thrombin, which in turn converts fibrinogen into fibrin (blood clots). FXa deficiency causes hemostasis defects, such as intracranial bleeding, hemathrosis, and gastrointestinal blood loss. He...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi500770p
更新日期：2014-11-11 00:00:00
abstract：:A method for the rapid and quantitative analysis of 5'-terminal oligonucleotides of RNAs made in vitro is described. The method involves synthesis of RNA in the presence of [gamma-32P]ATP or GTP, isolation of the RNA, and digestion with T1 or pancreatic ribonucleases to release labeled 5'-triphosphate termanated oligo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00604a005
更新日期：1978-05-30 00:00:00
abstract：:X-ray absorption spectroscopy at the Zn K-edge indicates that the active site of the marine diatom Thalassiosira weissflogii carbonic anhydrase is strikingly similar to that of mammalian alpha-carbonic anhydrase enzymes. The zinc has three histidine ligands and a single water at 1.98 A. This is quite different from th...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi001416s
更新日期：2000-10-10 00:00:00
abstract：:Helix-helix interactions within membranes are dominated by van der Waals packing motifs and side chain-side chain hydrogen bond formation, which act in tandem to determine the residues that comprise the interface between two given helices. To explore in a systematic manner the tertiary contacts between transmembrane h...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0492760
更新日期：2004-11-16 00:00:00
abstract：:Effects of different end sequences on melting, circular dichroism spectra (CD), and enzyme binding properties were investigated for four 40 base pair, non-self-complementary duplex DNA oligomers. The center sequences of these oligoduplexes have either of two 22 base pair modules flanked on both sides by sequences diff...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi000326k
更新日期：2000-07-04 00:00:00
abstract：:Human immunoglobulin E (IgE) consists of 14 domains, each with the characteristic immunoglobulin fold structure. Compared with the 12-domain structure of immunoglobulin G (IgG), IgE has an additional pair of domains (C epsilon 2) in the Fc region in place of the hinge of IgG. The crystal structure of the 4-domain Fc f...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00044a023
更新日期：1995-11-07 00:00:00
abstract：:The prothrombinase complex assembles through reversible interactions between factor Xa, factor Va and acidic phospholipid-containing membranes in the presence of calcium ions. This complex catalyses the conversion of prothrombin to thrombin through two proteolytic steps. We have used prethrombin 2 as a substrate analo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9623993
更新日期：1997-03-18 00:00:00
abstract：:Two dioxygenases (ARD and ARD') were cloned from Klebsiella pneumoniae that catalyze different oxidative decomposition reactions of an advanced aci-reductone intermediate, CH(3)SCH(2)CH(2)COCH(OH)=CH(OH) (I), in the methionine salvage pathway. The two enzymes are remarkable in that they have the same polypeptide seque...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi010110y
更新日期：2001-05-29 00:00:00
abstract：:It is shown that the protochlorophyllide ester pool of etiolated higher plants is a faithful copy of the protochlorophyllide pool. It is made up of both monovinyl- and divinylprotochlorophyllide esters. Although the two tetrapyrroles exhibited similar emission maxima, they were distinguishable by their Soret excitatio...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00562a027
更新日期：1980-10-14 00:00:00
abstract：:Endonuclease V from bacteriophage T4 is a well characterized enzyme that initiates the repair of ultraviolet light induced pyrimidine dimers. Scission of the phosphodiester backbone between the pyrimidines within a dimer, or 3' to an abasic (AP) site, occurs by a beta-elimination mechanism. In addition, high concentra...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00027a031
更新日期：1995-07-11 00:00:00
abstract：:Dihydroorotates deuteriated at both C5 and C6 have been prepared and used to probe the mechanism of the bovine liver mitochondrial dihydroorotate dehydrogenase. Primary deuterium isotope effects on kcat are observed with both (6RS)-[5(S)-2H]- and (6RS)-[6-2H] dihydroorotates (3 and 6, respectively); these effects are ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00429a041
更新日期：1989-02-07 00:00:00
abstract：:Homology modeling was used to identify two particular residues, Glu175 and Ala176, in Pseudomonas stutzeri phosphite dehydrogenase (PTDH) as the principal determinants of nicotinamide cofactor (NAD(+) and NADP(+)) specificity. Replacement of these two residues by site-directed mutagenesis with Ala175 and Arg176 both s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035018b
更新日期：2003-10-14 00:00:00
abstract：:G-Actin is a globular protein (Mr 42 300) known to have three cysteine residues that are at least partially exposed and chemically reactive (Cys-10, -284, and -374). When G-actin was reacted with 3-bromo-1,1,1-trifluoropropanone, three resolvable 19F resonances were observed in the 19F NMR spectrum. This fluorinated G...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00356a050
更新日期：1986-04-22 00:00:00
abstract：:In cerebellum, inositol trisphosphate- (InsP(3)-) gated Ca channels play a key role in learning, though they exhibit a low sensitivity to InsP(3) compared to peripheral tissues. In the present study, the cerebellar InsP(3) receptor is shown to be associated with a novel inhibitor of InsP(3) binding. (3)H-InsP(3) bindi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi992175v
更新日期：2000-03-28 00:00:00
abstract：:The synthesis of activated sulfate (adenosine 5'-phosphosulfate, APS) and inorganic pyrophosphate from ATP and SO4 is remarkably unfavorable: K(eq) approximately 10(-8) under presumed, near-physiological conditions. Consequently, ATP sulfurylases, which catalyze APS synthesis, suffer approximately 10(8)-fold losses in...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi060421e
更新日期：2006-09-26 00:00:00
abstract：:Tyrosine phenol-lyase, a tetrameric pyridoxal 5'-phosphate dependent enzyme, catalyzes the reversible hydrolytic cleavage of L-tyrosine to phenol and ammonium pyruvate. Here we describe the crystal structure of the Citrobacter freundii holoenzyme at 1.9 A resolution. The structure reveals a network of protein interact...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0601858
更新日期：2006-06-20 00:00:00
abstract：:Trypanosomes are purine-auxotrophic parasites that depend upon nucleoside hydrolase (NH) activity to salvage nitrogenous bases necessary for nucleic acid and cofactor synthesis. Nonspecific and purine-specific NHs have been widely studied, yet little is known about the 6-oxopurine-specific isozymes, although they are ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi100697d
更新日期：2010-10-19 00:00:00
abstract：:HbPresbyterian (beta 108Asn --> Lys, HbP) contains an additional positive charge (per alpha beta dimer) in the middle of the central cavity and exhibits a lower oxygen affinity than wild-type HbA in the presence of chloride. However, very little is known about the molecular origins of its altered functional properties...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9827464
更新日期：1999-08-31 00:00:00
abstract：:A cell-free system for synthesizing protein from wheat germ was used to translate the messenger RNA extracted from 16-day embryonic chick calvaria. A part of the product had properties similar to collagenous peptides and served as a substrate for prolyl hydroxylase, an enzyme specific for collagen. The level of potass...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00649a016
更新日期：1976-02-24 00:00:00
abstract：:Transport of protons and solutes across mitochondrial membranes is essential for many physiological processes. However, neither the proton-pumping respiratory chain complexes nor the mitochondrial secondary active solute transport proteins have been characterized electrophysiologically in their native environment. In ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1011755
更新日期：2010-12-07 00:00:00
abstract：:The crystal structure of the recombinant thiamin diphosphate-dependent E1 component from the Escherichia coli pyruvate dehydrogenase multienzyme complex (PDHc) has been determined at a resolution of 1.85 A. The E. coli PDHc E1 component E1p is a homodimeric enzyme and crystallizes with an intact dimer in an asymmetric...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0118557
更新日期：2002-04-23 00:00:00
abstract：:Phosphorus NMR spectroscopy is an important technique for the investigation of metabolism in tissues and intact organisms (including man). However, quantitation of the signals from an NMR experiment is difficult because it is not known from which regions of a cell metabolites are detected. It is generally believed tha...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00439a003
更新日期：1989-06-27 00:00:00
abstract：:Histone deacetylases have proven to be attractive novel targets for the treatment of cancer. The first inhibitor of histone deacetylases was approved for the treatment of cutaneous T-cell lymphoma in 2006. The identification of new lead structures with improved effectiveness and fewer side effects is necessary. This r...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi901617w
更新日期：2010-02-23 00:00:00
abstract：:Kinetic and structural changes in recombinant human aldose reductase (AR) due to modification by S-nitrosoglutathione (GSNO) were investigated. Incubation of the enzyme with 10-50 microM GSNO led to a time- and concentration-dependent inactivation of the enzyme, with a second-order rate constant of 0.087 +/- 0.009 M-1...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9714722
更新日期：1997-12-16 00:00:00
abstract：:Several polypeptide toxins from sea anemones caused delayed inactivation of mammalian voltage-dependent sodium channels, resulting in a positive inotropic effect on the heart. Anthopleurin B (ApB), a toxin produced by the sea anemone Anthopleura xanthogrammica, is the most potent of all known anemone toxins. Previous ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9528457
更新日期：1996-03-19 00:00:00
abstract：:Shaker potassium (K+) channels normally lack intrasubunit and intersubunit disulfide bonds. However, disulfide bonds are formed between Shaker subunits in intact cells exposed to oxidizing conditions. Upon electrophoresis under nonreducing conditions, intersubunit disulfide bond formation was detected by the presence ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi961083s
更新日期：1996-09-17 00:00:00
abstract：:Cytochrome P450 3A4 (CYP3A4) inhibitors ritonavir and cobicistat, currently administered to HIV patients as pharmacoenhancers, were designed on the basis of the chemical structure/activity relationships rather than the CYP3A4 crystal structure. To better understand the structural basis for CYP3A4 inhibition and the li...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi4005396
更新日期：2013-07-02 00:00:00
