Abstract:
:Electron spin echo envelope modulation (ESEEM) spectroscopy has been applied to the determination of the number of water molecules coordinated to the metal in the binary complex of staphylococcal nuclease with Mn2+, to the ternary enzyme-Mn2+-3',5'-pdTp complex, and to ternary complexes of a number of mutant enzymes in which metal-binding ligands have been individually altered. Quantitation of coordinated water is based on ESEEM spectral comparisons of Mn2+-EDTA and Mn2+-DTPA, which differ by a single inner sphere water, and with Mn2+-(H2O)6. It was found that Mn2+ in the ternary complex of the wild-type enzyme has a single additional coordinated water, as compared to Mn2+ in the binary complex, confirming earlier findings based on T1 measurements of bound water [Serpersu, E. H., Shortle, D. L., & Mildvan, A. S. (1987) Biochemistry 26, 1289-1300]. Ternary complexes of the mutant proteins D40E, D40G, and D21Y have the same number of water ligands as the ternary complex of the wild-type enzyme, while the D21E mutant has one less water ligand. In order to maintain octahedral coordination geometry, these findings require two additional ligands to Mn2+ from the protein in the binary complex of the wild-type enzyme, probably Glu 43 and Asp 19, and one additional ligand from the protein in the ternary D40G and D21E complexes. Other ESEEM studies of ternary Mn2+ complexes of wild-type, D21E, and D21Y mutants indicate the coordination by Mn2+ of a phosphate of 3',5'-pdTp, as demonstrated by a 31P contact interaction of 3.9 +/- 0.3 MHz. Magnetic interaction of Mn2+ with 31P could not be demonstrated with the D40G and D40E mutants, suggesting that metal-phosphate distances are greater in these mutants than in the wild-type protein. In a parallel NMR study of the paramagnetic effects of enzyme-bound Co2+ (which occupies the Mn2+ site on the enzyme) on the T1 of 31P from enzyme-bound 3',5'-pdTp and 5'-TMP, it was found that metal to 5'-phosphate distances are 0.9-1.6 A shorter in ternary complexes of the wild-type enzyme and of the D21E mutant than in ternary complexes of the D40G mutant. In all cases, the 5'-phosphate of pdTp is in the inner coordination sphere of Co2+ and the 3'-phosphate is predominantly in the second coordination sphere.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Serpersu EH,McCracken J,Peisach J,Mildvan ASdoi
10.1021/bi00421a010subject
Has Abstractpub_date
1988-10-18 00:00:00pages
8034-44issue
21eissn
0006-2960issn
1520-4995journal_volume
27pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::We have studied the hydrogen-exchange kinetics of hemoglobin A0 as a function of ligand, CO, saturation. In the noncooperative system, azide binding to methemoglobin, the alterations in exchange kinetics are proportional to the average degree of ligation. However, in the case of CO binding to deoxyhemoglobin the chang...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00297a015
更新日期:1984-01-17 00:00:00
abstract::We constructed and expressed chimeric receptor cDNAs with insulin receptor exon 3 (residues 191-297 of the cysteine-rich region) replaced with either the comparable region of the insulin-like growth factor I receptor (IGF-IR) or the insulin receptor related receptor (IRR). Both chimeric receptors still could bind insu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00235a001
更新日期:1991-05-28 00:00:00
abstract::The contact between EcoRI endonuclease and the "primary clamp" phosphate of its recognition site pGAATTC is absolutely required for recognition of the canonical and all variant DNA sites. We have probed this contact using oligonucleotides containing single stereospecific (Rp)- or (Sp)- phosphorothioates (Ps). At the G...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960261e
更新日期:1996-07-09 00:00:00
abstract::Crk-II is a signaling adaptor protein that is involved in many cellular processes including apoptosis, proliferation, and differentiation. It has a modular domain architecture consisting of an Src homology 2 domain (SH2) followed by two Src homology 3 (SH3) domains. The structures and ligand-binding properties of the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060590z
更新日期:2006-07-25 00:00:00
abstract::The family 4 uracil-DNA glycosylase from the hyperthermophilic organism Archaeoglobus fulgidus (AFUDG) is responsible for the removal of uracil in DNA as the first step in the base excision repair (BER) pathway. AFUDG contains a large solvent-exposed peptide region containing an α helix and loop anchored on each end v...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi3000462
更新日期:2012-06-26 00:00:00
abstract::There are at least nine neurodegenerative diseases associated with proteins that contain an unusually expanded polyglutamine domain, the best known of which is Huntington's disease. In all of these diseases, the mutant protein aggregates into neuronal inclusions; it is generally, although not universally, believed tha...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700806c
更新日期:2007-11-06 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::Gramicidin crystals, dimyristoylphosphatidylcholine (DMPC)/gramicidin dispersions, and DMPC/gramicidin cocrystals were examined by Raman scattering to determine lipid/gramicidin stoichiometries and lipid organization. Calibrations of the choline (716-cm-1) and tryptophan (756-cm-1) peaks indicate that the cocrystals c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00376a030
更新日期:1987-01-27 00:00:00
abstract::Catalase HPII from Escherichia coli is a homotetramer of 753 residue subunits. The multimer displays a number of unusual structural features, including interwoven subunits and a covalent bond between Tyr415 and His392, that would contribute to its rigidity and stability. As the temperature of a solution of HPII in 50 ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982863z
更新日期:1999-03-30 00:00:00
abstract::The interaction of nile red (NR) with apomyoglobin (ApoMb) in the native (pH 7) and molten globule (pH 4) states was investigated using experimental and computational methods. NR binds to hydrophobic locations in ApoMb with higher affinity (K(d) = 25 +/- 5 microM) in the native state than in the molten globule state (...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051905y
更新日期:2006-04-25 00:00:00
abstract::The MexA-MexB-OprM efflux pump exports structurally and functionally diverse xenobiotics and confers multi-drug resistance on Pseudomonas aeruginosa cells. The MexB transporter traverses the inner membrane twelve times, bears two large periplasmic domains and has two homologous tandem repeats. To test whether two homo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0300074
更新日期:2003-06-17 00:00:00
abstract::Phospholipase B1 (PLB1), secreted by the pathogenic yeast Cryptococcus neoformans, has an established role in virulence. Although the mechanism of its phospholipase B, lysophospholipase, and lysophospholipase transacylase activities is unknown, it possesses lipase, subtilisin protease aspartate, and phospholipase moti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7009508
更新日期:2007-09-04 00:00:00
abstract::PLK1 (polo-like kinase 1) is a key mitotic kinase and a therapeutic target in the treatment of proliferative diseases. Here we investigate the relative substrate specificity and pharmacological relatedness of PLK1, -2, -3, and -4 that together comprise a conserved family of Ser/Thr kinases (PLK family). We report cons...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7008745
更新日期:2007-08-21 00:00:00
abstract::Genetic knockout of the BtR4 gene encoding the Heliothis virescens cadherin-like protein (HevCaLP) is linked to resistance against Cry1Ac toxin from Bacillus thuringiensis. However, the functional Cry1Ac receptor role of this protein has not been established. We previously proposed HevCaLP as a shared binding site for...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0606703
更新日期:2006-08-15 00:00:00
abstract::Adenosine 5'-phosphosulfate (APS) kinase catalyzes the second reaction in the two-step conversion of inorganic sulfate to 3'-phosphoadenosine 5'-phosphosulfate (PAPS). This report presents the 2.0 A resolution crystal structure of ligand-free APS kinase from the filamentous fungus, Penicillium chrysogenum. The enzyme ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9924157
更新日期:2000-02-22 00:00:00
abstract::Potent inhibitors of human cysteine proteases of the papain family have been made and assayed versus a number of relevant family members. We describe the synthesis of peptide alpha-ketoheterocyclic inhibitors that occupy binding subsites S1'-S3 of the cysteine protease substrate recognition cleft and that form a rever...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035041x
更新日期:2003-12-30 00:00:00
abstract::Green fluorescent protein and its variants are frequently used as Förster (fluorescence) resonance energy transfer (FRET) pairs to determine the proximity of protein domains. We prepared fusion proteins comprising yellow fluorescent protein-Dictyostelium myosin II motor domain-cyan fluorescent protein (YFP-myosin-CFP)...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060943u
更新日期:2006-09-05 00:00:00
abstract::Multiply branched polylactosaminoglycans are expressed in glycoproteins and glycolipids of many cells. Interest in their biology stems from their abundant expression in early embryonal cells and from their ability to carry multiple lectin-binding determinants, which makes them prominent ligands and antagonists of cell...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9627673
更新日期:1997-06-10 00:00:00
abstract::Protein kinase C (PKC) interacted with phospholipid vesicles in a calcium-dependent manner and produced two forms of membrane-associated PKC: a reversibly bound form and a membrane-inserted form. The two forms of PKC were isolated and compared with respect to enzyme stability, cofactor requirements, and phorbol ester ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00420a003
更新日期:1988-10-04 00:00:00
abstract::The selenoamino acids methylselenocysteine (MeSeCys) and selenomethionine (SeMet) have disparate efficacies as anticancer agents. Herein, we use X-ray absorption spectroscopy to determine the chemical form of selenium in human neuroblastoma cells. Cells treated with MeSeCys contain a significant diselenide component, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201462u
更新日期:2012-01-24 00:00:00
abstract::The properties of a series of new potentiometric membrane probes have been explored. The probes all contain an (aminostyryl)pyridinium chromophore or a more highly conjugated analogue. The spectral properties of the dyes are discussed in terms of the excitation-induced charge shift from the pyridine to the aniline; th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00342a010
更新日期:1985-10-08 00:00:00
abstract::Eosinophil cationic protein (ECP) is a component of the eosinophil granule matrix. It shows marked toxicity against helminth parasites, bacteria single-stranded RNA viruses, and host epithelial cells. Secretion of human ECP is related to eosinophil-associated allergic, asthmatic, and inflammatory diseases. ECP belongs...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0264521
更新日期:2002-10-08 00:00:00
abstract::The binding of 125I-labeled ovine follitropin (oFSH) and 125I-labeled deglycosylated ovine follitropin (DG-oFSH) to porcine granulosa cell membranes was studied at equilibrium and nonequilibrium binding conditions and statistically analyzed. Saturation and competition binding experiments revealed homogeneity in the po...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00386a059
更新日期:1987-06-16 00:00:00
abstract::myo-Inositol (mI) is widely distributed in all domains of life and is important for several cellular functions, including bacterial survival. The enzymes responsible for the bacterial catabolism of mI, encoded in the iol operon, can vary from one organism to another, and these pathways have yet to be fully characteriz...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00539
更新日期:2020-08-18 00:00:00
abstract::Azurin from Pseudomonas aeruginosa and two mutants where the methionine ligand has been mutated have been studied in order to directly investigate the functional and structural significance of this ligand in the blue copper proteins. Reduction potentials, X-ray absorption fine structure (XAFS), electron paramagnetic r...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00059a013
更新日期:1993-03-02 00:00:00
abstract::Protein amyloid fibrils are originally identified as pathological entities in a variety of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Recent studies have revealed that amyloid fibrils also serve as functional protein assemblies to fulfill a wide range of biological functions. Decip...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/acs.biochem.9b01069
更新日期:2020-02-11 00:00:00
abstract::Most of the molybdoenzymes contain, in the amino-terminal region of their catalytic subunits, a conserved Cys group that in some cases binds an [Fe-S] cluster. In dissimilatory nitrate reductases, the first Cys residue of this motif is replaced by a conserved His residue. Site-directed mutagenesis of this residue (His...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972858f
更新日期:1998-05-19 00:00:00
abstract::Sedimentation in a preparative ultracentrifuge was used to determine the affinity of heavy meromyosin, HMM, for regulated actin, F-actin plus troponin-tropomyosin, in the presence of MgATP at pH 7.0, 20 degrees C, and mu = 18 mM. HMM was prepared from vertebrate striated muscle myosin by a mild chymotryptic digestion....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00275a003
更新日期:1983-03-15 00:00:00
abstract::The heme-regulated inhibitor (HRI) of protein synthesis becomes activated in rabbit reticulocyte lysates in response to a variety of conditions including heme-deficiency, addition of oxidants, and heat shock. Activated HRI inhibits translation by catalyzing the phosphorylation of the alpha-subunit of eukaryotic initia...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00080a001
更新日期:1993-07-27 00:00:00
abstract::MT1-MMP, a prototypic member of a membrane-type metalloproteinase subfamily, is an invasion promoting protease and an activator of MMP-2. In addition, MT1-MMP proteolysis regulates the functionality of cell-surface adhesion/signaling receptors including tissue transglutaminase (tTG). tTG is known to serve as an adhesi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049266z
更新日期:2004-09-21 00:00:00