Abstract：
:The local environment of the transmembrane and C-terminal domain of M13 major coat protein was probed by site-directed ESR spin labeling when the protein was introduced into three membrane-mimicking systems, DOPC vesicles, sodium cholate micelles, and SDS micelles. For this purpose, we have inserted unique cysteine residues at specific positions in the transmembrane and C-terminal region, using site-directed mutagenesis. Seven viable mutants with reasonable yield were harvested: A25C, V31C, T36C, G38C, T46C, A49C, and S50C. The mutant coat proteins were indistinguishable from wild type M13 coat protein with respect to their conformational and aggregational properties. The ESR data suggest that the amino acid positions 25 and 46 of the coat protein in DOPC vesicles are located close to the membrane-water interface. In this way the lysines at positions 40, 43, and 44 and the phenylalanines at positions 42 and 45 act as hydrophilic and hydrophobic anchors, respectively. The ESR spectra of site specific maleimido spin-labeled mutant coat proteins reconstituted into DOPC vesicles and solubilized in sodium cholate or SDS indicate that the local dynamics of the major coat protein is significantly affected by its structural environment (micellar vs bilayer), location (aqueous vs hydrophobic), and lipid/protein ratio. The detergents SDS and sodium cholate sufficiently well solubilize the major coat protein and largely retain its secondary structure elements. However, the results indicate that they have a poorly defined protein-amphiphilic structure and lipid-water interface as compared to bilayers and thus are not a good substitute for lipid bilayers in biophysical studies.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Stopar D,Spruijt RB,Wolfs CJ,Hemminga MAdoi
10.1021/bi961770jsubject
Has Abstractpub_date
1996-12-03 00:00:00pages
15467-73issue
48eissn
0006-2960issn
1520-4995pii
bi961770jjournal_volume
35pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Spatially localized differences in the conformational dynamics of the rubredoxins from the hyperthermophile Pyrococcus furiosus (Pf) and the mesophile Clostridium pasteurianum (Cp) are monitored via amide exchange measurements. As shown previously for the hyperthermophile protein, nearly all backbone amides of the Cp ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0112560
更新日期：2001-12-04 00:00:00
abstract：:Crude calf thymus DNA-dependent RNA polymerase, RNA polymerase B (ribonucleoside triphosphate: RNA nucleotidyltransferase, EC 2.7.7.6), was incubated with the tritium labeled, potent inhibitor [3H]amanin, in order to form the enzymatically inactive [3H]amanin-polymerase complex ([3H]A-P complex). Subsequent purificati...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00661a013
更新日期：1976-08-10 00:00:00
abstract：:Adenosine cyclic 3',5'-phosphate receptor protein (CRP or CAP) is a regulatory protein involved in the transcription of several operons in Escherichia coli. cAMP-independent, nonspecific complexes of CRP and DNA were investigated by photochemical cross-linking of the protein to nonspecific DNA, whose thymines are subs...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00058a010
更新日期：1993-02-23 00:00:00
abstract：:The chirality of sulfoxidation catalyzed by two cytochrome P-450 isozymes purified from phenobarbital-induced rat liver was studied by using 4-tolyl ethyl sulfide as a substrate. Both P-450 isoenzymes, termed PB-1 and PB-4, when reconstituted with purified rat liver NADPH-cytochrome P-450 reductase and cytochrome b5, ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00539a032
更新日期：1982-05-11 00:00:00
abstract：:The proton nuclear magnetic resonance spectrum of human adult deoxyhemoglobin in D2O in the region from 6 to 20 ppm downfield from the proton resonance of residual water shows a number of hyperfine shifted proton resonances that are due to groups on or near the alpha and beta hemes. The sensitivity of these resonances...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00590a031
更新日期：1979-11-13 00:00:00
abstract：:Tryptophan 2,3-dioxygenase (TDO) from Xanthomonas campestris is a highly specific heme-containing enzyme from a small family of homologous enzymes, which includes indoleamine 2,3-dioxygenase (IDO). The structure of wild type (WT TDO) in the catalytically active, ferrous (Fe (2+)) form and in complex with its substrate...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi801202a
更新日期：2008-10-07 00:00:00
abstract：:As part of a search for organic compounds that selectively target RNA, we found that specific diphenylfuran derivatives, which are related to compounds that bind to the DNA minor groove, bind very strongly to RNA in a manner very sensitive to the structure of the compounds. In extended development of the diphenylfuran...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi002338b
更新日期：2001-02-06 00:00:00
abstract：:The heterotrimeric G protein subunit, alpha(s), can move reversibly from plasma membranes to cytoplasm in response to activation by GPCRs or activating mutations. We examined the importance of the unique N-terminus of alpha(s) in this translocation in cultured cells. alpha(s) contains a single site for palmitoylation ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi025533u
更新日期：2002-07-30 00:00:00
abstract：:Phospholipase A(2) (PLA(2)) enzymes become activated by binding to biological membranes and hydrolyze phospholipids to free fatty acids and lyso-phospholipids, the precursors of inflammatory mediators. To understand the functional significance of amino acid residues at key positions, we have studied the effects of the...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi061440r
更新日期：2006-10-17 00:00:00
abstract：:The first positive evidence for the utilization of a direct C-6' ' oxidation/reduction mechanism by ADP-l-glycero-d-manno-heptose 6-epimerase is reported here. The epimerase (HldD or AGME, formerly RfaD) operates in the biosynthetic pathway of l-glycero-d-manno-heptose, which is a conserved sugar in the core region of...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi050106c
更新日期：2005-04-19 00:00:00
abstract：:Addition of short sequences of dCMP residues to the 3'-OH end of duplex linear DNAs allows rapid and efficient transcription to be initiated at these sites by purified mammalian RNA polymerase II [Kadesch, T. R., & Chamberlin, M. J. (1982) J. Biol. Chem. 257, 5286-5295]. The use of such tailed DNA templates should all...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00330a019
更新日期：1985-04-23 00:00:00
abstract：:The thermodynamics of L-tryptophan and operator DNA binding to the tryptophan repressor of Escherichia coli were analyzed by titration microcalorimetry and van't Hoff analysis of footprinting titrations, respectively. At 25 degrees C in 10 mM sodium phosphate, pH 7.6, and 0.1 M NaCl, the binding of L-tryptophan to the...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00079a029
更新日期：1993-07-20 00:00:00
abstract：:The guanidine hydrochloride- (GuHCl-) induced unfolding and refolding of a recombinant domain of bovine microsomal cytochrome b(5) containing the first 104 amino acid residues has been characterized by both transient and equilibrium spectrophotometric methods. The soluble domain is reversibly unfolded and the equilibr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi991413j
更新日期：1999-10-26 00:00:00
abstract：:Synthesis of a novel perylene-neomycin conjugate (3) and the properties of its binding to human telomeric G-quadruplex DNA, 5'-d[AG3(T2AG3)3] (4), are reported. Various spectroscopic techniques were employed to characterize the binding of conjugate 3 to 4. A competition dialysis assay revealed that 3 preferentially bi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1017304
更新日期：2011-04-12 00:00:00
abstract：:The kinetics and thermodynamics of Al(III) exchange between aluminum citrate (AlL) and human serum transferrin were investigated in the 7.2-8.9 pH range. The C-site of human serum apotransferrin in interaction with bicarbonate removes Al(III) from Al citrate with an exchange equilibrium constant K1 = (2.0 +/- 0.6) x 1...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi020627p
更新日期：2003-03-18 00:00:00
abstract：:Preprotein translocation in Escherichia coli is mediated by the translocase with SecA as peripheral ATPase and SecY, SecE, and SecG as membrane domain. To facilitate large-scale purification of the SecYEG heterotrimer, SecY was fused at its amino terminus with a hexahistidine tag and co-overexpressed with SecE and Sec...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi972105t
更新日期：1998-01-06 00:00:00
abstract：:Despite much work, subcellular neurons of Caenorhabditis elegans have not been studied at nanometer resolution with millisecond time resolution. Nor has there been an effective way to immobilize C. elegans. Here we show that, without using anesthetic or paralyzing agents, fluorescence imaging with one-nanometer accura...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8023477
更新日期：2009-06-09 00:00:00
abstract：:A mass transport theory of the gel chromatographic procedure of Hummel and Dreyer for measuring the binding of small ligand molecules to macromolecules has been formulated for ligand-mediated macromolecular association. It is concluded that the Hummel-Dreyer procedure is the method of choice for quantitating ligand bi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00666a011
更新日期：1976-10-19 00:00:00
abstract：:The repair of the endogenous lesion 8-oxo-7,8-dihydrodeoxyguanosine (8-oxodG) was investigated in the nucleotide excision repair mutant xeroderma pigmentosum D (XPD), using human normal or transformed XPD fibroblasts and the Chinese hamster XPD cell line UV5. In vivo repair of 8-oxodG induced by hydrogen peroxide trea...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi992610l
更新日期：2000-08-29 00:00:00
abstract：:Myocardial contraction is initiated when Ca2+ binds to site II of cardiac troponin C. This 12-residue EF-hand loop (NH2-DEDGSGTVDFDE-COOH) contains six residues (bold) that coordinate Ca2+ binding and six residues that do not appear to influence Ca2+ binding directly. We have introduced six single-cysteine substitutio...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi800164c
更新日期：2008-05-06 00:00:00
abstract：:A series of truncated forms of His(6)-tagged gp91phox were expressed, solubilized, and purified in the presence of 30 microM FAD. The truncated gp91phox with the longest sequence in the C-terminal region (221-570) (gp91C) showed the highest activity (turnover rate, 0.92) for NADPH diaphorase in the presence of either ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0400249
更新日期：2004-07-27 00:00:00
abstract：:6-Methylsalicylic acid synthase, the multifunctional enzyme complex that catalyzes the biosynthesis of the tetraketide 6-methylsalicylic acid, was modified by thiol-specific inhibitors and cross-linking reagents. Treatment with 1,3-dibromopropan-2-one caused rapid enzyme inactivation and formation of cross-linked dime...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960422e
更新日期：1996-09-24 00:00:00
abstract：:From a series of 15N-resolved 3D ROESY-HMQC and 13C-resolved 3D NOESY-HMQC spectra of the FK506 binding protein (FKBP)/ascomycin complex in H2O, the locations of three tightly bound water molecules were identified. These waters are all buried within the interior of the complex and form an integral part of its structur...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00061a004
更新日期：1993-03-16 00:00:00
abstract：:Phospholipids containing a saturated fatty acid in the primary position and an unsaturated fatty acid in the secondary position are a major structural part of biological membranes. The mixed-chain hydrophobic core of the membranes is the diacylglycerol part. To better understand the core properties of membranes we hav...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00051a015
更新日期：1995-12-26 00:00:00
abstract：:Carbon monoxide dehydrogenase (CODH) from Clostridium thermoaceticum plays a central role in the newly discovered acetyl-CoA pathway [Wood, H.G., Ragsdale, S.W., & Pezacka, E. (1986) FEMS Microbiol. Rev. 39, 345-362]. The enzyme catalyzes the formation of acetyl-CoA from methyl, carbonyl, and CoA groups, and it has sp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00417a045
更新日期：1988-08-23 00:00:00
abstract：:Porcine aortic myosin is a smooth muscle contractile protein similar to its striated muscle counterpart. Electrophoresis in sodium dodecyl sulfate indicates that the molecule consists of three classes of subunits with polypeptide chain molecular weights of 192,000, 19,000, and 15,000. At 277 nm the absorption spectrum...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00576a003
更新日期：1979-05-01 00:00:00
abstract：:The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 binding. The crystal structure of the B...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi801115g
更新日期：2008-11-04 00:00:00
abstract：:Members of the ATP-binding cassette (ABC) transporters share the same basic architecture, with a four-core domain made of two transmembrane plus two nucleotide-binding domains. However, a supramolecular organization has been detected in some ABC transporters, which might be relevant to physiological regulation of subs...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0482809
更新日期：2005-03-22 00:00:00
abstract：:6-[(4-Bromo-2,3-dioxobutyl)thio]-6-deaminoadenosine 5'-diphosphate (6-BDB-TADP) has been shown to react at the reduced diphosphopyridine nucleotide (DPNH) inhibitory site of bovine liver glutamate dehydrogenase with incorporation of 1 mol of reagent/mol of enzyme subunit [Batra, S. P., & Colman, R. F. (1984) Biochemis...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00360a005
更新日期：1986-06-17 00:00:00
abstract：:The human intestinal Caco-2 cell line, described as enterocyte-like in a number of studies, was examined for its ability to carry out the metabolism of vitamin A normally required in the absorptive process. Caco-2 cells contained cellular retinol-binding protein II, a protein which is abundant in human villus-associat...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00502a015
更新日期：1990-12-18 00:00:00
