Abstract：
:Enzyme therapeutics that can degrade l-methionine (l-Met) are of great interest as numerous malignancies are exquisitely sensitive to l-Met depletion. To exhaust the pool of methionine in human serum, we previously engineered an l-Met-degrading enzyme based on the human cystathionine-γ-lyase scaffold (hCGL-NLV) to circumvent immunogenicity and stability issues observed in the preclinical application of bacterially derived methionine-γ-lyases. To gain further insights into the structure-activity relationships governing the chemistry of the hCGL-NLV lead molecule, we undertook a biophysical characterization campaign that captured crystal structures (2.2 Å) of hCGL-NLV with distinct reaction intermediates, including internal aldimine, substrate-bound, gem-diamine, and external aldimine forms. Curiously, an alternate form of hCGL-NLV that crystallized under higher-salt conditions revealed a locally unfolded active site, correlating with inhibition of activity as a function of ionic strength. Subsequent mutational and kinetic experiments pinpointed that a salt bridge between the phosphate of the essential cofactor pyridoxal 5'-phosphate (PLP) and residue R62 plays an important role in catalyzing β- and γ-eliminations. Our study suggests that solvent ions such as NaCl disrupt electrostatic interactions between R62 and PLP, decreasing catalytic efficiency.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Yan W,Stone E,Zhang YJdoi
10.1021/acs.biochem.6b01172subject
Has Abstractpub_date
2017-02-14 00:00:00pages
876-885issue
6eissn
0006-2960issn
1520-4995pii
10.1021/acs.biochem.6b01172journal_volume
56pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:The photocycle of the blue-light photoreceptor protein Photoactive Yellow Protein (PYP) was studied at reduced relative humidity (RH). Photocycle kinetics and spectra were measured in thin films of PYP in which the relative humidity was set at values between 29 and 98% RH with saturated solutions of various salts. We ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi050237d
更新日期：2005-06-28 00:00:00
abstract：:The intense charge transfer complex between the enolate of 3-thia-octanoyl-CoA and the oxidized flavin of the medium-chain acyl-CoA dehydrogenase is discharged by the ferricenium ion with irreversible inactivation of the enzyme. Charge transfer complex formation is a necessary, but insufficient, condition for oxidativ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00050a025
更新日期：1995-12-19 00:00:00
abstract：:The specificity of trypsin for Arg- and Lys-containing substrates depends upon the presence of Asp189 at the base of the primary binding pocket. The crystal structure of anionic rat trypsin D189S complexed with BPTI reveals that removal of the aspartate side chain permits the binding of a well-ordered acetate ion in a...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00177a016
更新日期：1994-03-22 00:00:00
abstract：:A molecular model has been built of the equine infectious anemia virus (EIAV) proteinase on the basis of the crystal structures of the related Rous sarcoma virus (RSV) and human immunodeficiency virus (HIV) proteinases. The 104 residue long EIAV proteinase has 30 identical and 11 similar amino acids compared to those ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00064a019
更新日期：1993-04-06 00:00:00
abstract：:The binding of 8-anilino-1-naphthalenesulfonate (ANS) to ciliary dynein ATPase leads to a marked increase in the dye's fluorescence intensity, accompanied by a blue shift in the observed fluorescence emission maximum. We found that dynein has 37 +/- 3 ANS binding sites and that experimentally applied ANS concentration...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00347a012
更新日期：1985-12-17 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：:A grand canonical formalism is developed to combine discrete simulations for chemically distinct species in equilibrium. Each simulation is based on a perturbed funneled landscape. The formalism is illustrated using the alkaline-induced transitions of cytochrome c as observed by FTIR spectroscopy and with various othe...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi802293m
更新日期：2009-03-24 00:00:00
abstract：:The mechanism of oxidation of a peptide substrate by the flavoprotein lysine-specific demethylase (LSD1) has been examined using the effects of pH and isotopic substitution on steady-state and rapid-reaction kinetic parameters. The substrate contained the 21 N-terminal residues of histone H3, with a dimethylated lysyl...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi900499w
更新日期：2009-06-16 00:00:00
abstract：:The properties of neuronal nitric oxide synthase containing one tetrahydrobiopterin (BH4) per dimer [nNOS(BH4+)] were compared to those of the BH4-free enzyme [nNOS(BH4-)]. The stimulation by BH4 of the formation of L-citrulline at the expense of H2O2 production unambiguously demonstrated that BH4 is essential in coup...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi961931j
更新日期：1996-12-24 00:00:00
abstract：:A full-length cDNA for dopamine beta-monooxygenase (D beta M) from bovine adrenal glands has been cloned and sequenced. The soluble and membrane-derived forms of D beta M have also been sequenced from their N-termini. While the observed sequences for the soluble protein correspond to those previously reported [Joh, T....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00452a026
更新日期：1989-12-26 00:00:00
abstract：:The interactions of 4-thiouridine and 5-[(methylamino)methyl]-2-thiouridine in tRNA and of phosphorothioate esters in nucleic acids with an organomercurial have been investigated. For this purpose, an affinity electrophoretic system has been developed in which the mercury derivative has been covalently immobilized in ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00410a048
更新日期：1988-05-17 00:00:00
abstract：:FLI1 (Friend leukemia integration 1) is a metazoan transcription factor that is upregulated in a number of cancers. In addition, rearrangements of the fli1 gene cause sarcomas, leukemias, and lymphomas. These rearrangements encode oncogenic transcription factors, in which the DNA binding domain (DBD or ETS domain) of ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b01121
更新日期：2015-12-22 00:00:00
abstract：:The requirement for Ca2+ in the Mn(2+)-dependent photoactivation of oxygen evolution was re-evaluated using 17 kDa/24 kDa-less photosystem II (PSII) membranes depleted of (Mn)4-clusters by NH2OH extraction. At optimum conditions (1 mM Mn2+/10 microM 2,6-dichlorphenolindophenol (DCIP)/20 mM Ca2+), the light-induced inc...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00041a031
更新日期：1995-10-17 00:00:00
abstract：:The respiratory chain of Vibrio cholerae contains three bd-type quinol oxygen reductases as well as one cbb(3) oxygen reductase. The cbb(3) oxygen reductase has been previously isolated and characterized; however, the natural mobile electron donor(s) that shuttles electrons between the bc(1) complex and the cbb(3) oxy...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1004574
更新日期：2010-09-07 00:00:00
abstract：:The transformation of DMPC vesicles containing myristic acid to larger structures was examined by nuclear magnetic resonance techniques. The process was found to be characterized by a rate which is approximately first order in vesicle concentration. Rates, however, show a dramatic increase with increasing myristic aci...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00610a027
更新日期：1978-08-22 00:00:00
abstract：:Histone deacetylase 8 (HDAC8) is a well-characterized member of the class I acetyl-lysine deacetylase (HDAC) family. Previous work has shown that the efficiency of HDAC8-catalyzed deacetylation of a methylcoumarin peptide varies depending on the identity of the divalent metal ion in the HDAC8 active site. Here we demo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00851
更新日期：2017-10-24 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：:Adenylosuccinate lyase of Bacillus subtilis is inactivated by 2-[(4-bromo-2,3-dioxobutyl)thio]adenosine 5'-monophosphate (2-BDB-TAMP) at pH 7.0. As the reagent concentration is increased, a maximum rate constant is approached, indicative of reversible enzyme-reagent complex formation (KR = 68 +/- 9 microM) prior to ir...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9805339
更新日期：1998-06-09 00:00:00
abstract：:The present study uses CO as a surrogate for oxygen to probe how substrate binding triggers oxygen activation in peptidylglycine monooygenase (PHM). Infrared stretching frequencies (ν(C ≡ O)) of the carbonyl (CO) adducts of copper proteins are sensitive markers of Cu(I) coordination and are useful in probing oxygen re...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.6b00845
更新日期：2016-12-06 00:00:00
abstract：:Apo horse spleen ferritin (apo HoSF) was reconstituted to various core sizes (100-3500 Fe3+/HoSF) by depositing Fe(OH)3 within the hollow HoSF interior by air oxidation of Fe2+. Fe2+ and phosphate (Pi) were then added anaerobically at a 1:4 ratio, and both Fe2+ and Pi were incorporated into the HoSF cores. The resulti...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi982727u
更新日期：1999-05-18 00:00:00
abstract：:One of the distinctive features of eubacterial retinal-based proton pumps, proteorhodopsins, xanthorhodopsin, and others, is hydrogen bonding of the key aspartate residue, the counterion to the retinal Schiff base, to a histidine. We describe properties of the recently found eubacterium proton pump from Exiguobacteriu...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi300409m
更新日期：2012-07-24 00:00:00
abstract：:Administration of pregnenolone-16 alpha-carbonitrile (PCN) to adult female rats caused a 2-fold increase in total liver microsomal cytochrome P-450 along with 5-7-fold increases in four in vitro monooxygenase activities considered diagnostic for the major PCN-inducible cytochrome P-450 isozyme. However, upon administr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00387a022
更新日期：1987-06-30 00:00:00
abstract：:The inhibitory effects of urea on the normally rapid fibrillation of human calcitonin (hCT) were investigated by 1H NMR. From subtle differences in the chemical shift of hCT in the presence and absence of urea, the occurrence of weak interactions between urea and hCT was confirmed. The chemical shifts on the NH and C ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi961013l
更新日期：1996-10-01 00:00:00
abstract：:Human hephaestin (Hp) is a transmembrane protein that has been implicated in duodenal iron export. Mutations in the murine hephaestin gene (sla) produce microcytic, hypochromic anemia that is refractory to oral iron therapy. Hp shares approximately 50% sequence identity with the plasma multicopper ferroxidase cerulopl...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051559k
更新日期：2005-11-15 00:00:00
abstract：:We have studied the structure of the CuB site in the binuclear heme-copper center of the fully oxidized form of the quinol-oxidizing cytochrome aa3-600 from Bacillus subtilis by EXAFS and ENDOR spectroscopy. This enzyme is member of the large superfamily of heme-copper respiratory oxidases, which catalyze the reductio...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00032a019
更新日期：1995-08-15 00:00:00
abstract：:A cDNA coding for a P450 expressed in human lung was isolated from a lambda gt11 library constructed from human lung mRNA using a cDNA probe to rat P450 IVA1. The cDNA-deduced amino acid sequence of this P450, designated IVB1, consisted of 511 amino acids and had a calculated molecular weight of 59,558. The IVB1 amino...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00446a014
更新日期：1989-10-03 00:00:00
abstract：:The Vmax/Km (microM -1 min -1.) for bovine factor X activation by bovine factor IXa alpha, in the presence of sufficient [Ca2+] to saturate the initial reaction rate, was 0.007. When factor IXa beta was substituted for factor IXa alpha in this reaction, the Vmax/Km decreased to 0.001, suggesting that factor IXa alpha ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00286a007
更新日期：1983-08-16 00:00:00
abstract：:The coordination complex tris(4,7-diphenyl-1,10-phenanthroline)rhodium(III) [Rh(DIP)3(3+)], which promotes RNA cleavage upon photoactivation, has been shown to target specifically guanine-uracil (G-U) mismatches in double-helical regions of folded RNAs. Photoactivated cleavage by Rh(DIP)3(3+) has been examined on a se...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00139a001
更新日期：1992-06-23 00:00:00
abstract：:The structure of the His-94-->Cys variant of human carbonic anhydrase II (CAII) has been determined by X-ray crystallographic methods to a resolution of 2.3 A with a final crystallographic R factor of 0.155. This variant of CAII crystallizes in orthorhombic space group P2(1)2(1)2(1) which is the first example of a new...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00057a015
更新日期：1993-02-16 00:00:00
abstract：:Human erythrocyte band 3 inhibits glycolytic enzymes, including aldolase, by binding these cytoplasmic enzymes at its N-terminus. Phosphorylation of Y8 disrupts inhibition, and there is evidence that in vivo glycolysis levels in erythrocytes are regulated in part by a phosphorylation/dephosphorylation signaling pathwa...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00051a005
更新日期：1995-12-26 00:00:00
