Abstract:
:Conformational alterations occurring in bovine alpha 2-macroglobulin (alpha 2M) resulting from proteolysis and nucleophilic modification have been monitored by UV difference spectra, circular dichroism, and changes in the fluorescence of 6-(p-toluidino)-2-naphthalenesulfonate (TNS) and bis(8-anilino-1-naphthalenesulfonate) (Bis-ANS). The results of this study indicate that these two dyes appear capable of differentiating between conformational changes induced by proteolysis and those induced by methylamine treatment. It appears that TNS is a sensitive probe for monitoring protease-induced but not methylamine-induced conformational changes in bovine alpha 2M. Bis-ANS, on the other hand, appears suitable for monitoring conformational changes induced by methylamine treatment or proteolysis of the molecule and was used as a probe to monitor the kinetics of the conformational change induced by methylamine treatment. It was found that the conformational change did not occur simultaneously with cleavage of the thiol ester bonds by the nucleophile, measured by titration of free sulfhydryl groups with 5,5'-dithiobis(2-nitrobenzoate). The data are consistent with a model in which initial nucleophilic attack results in exposure of sulfhydryl groups, resulting in a conformational change measured by an increase in fluorescence. This event is followed by a unimolecular step representing a conformational change in the protein that results in a further increase in the fluorescence signal. The second-order rate constant for hydrolysis of the thiol ester bonds was determined to be 3.4 +/- 1.0 M-1 s-1, while the rate constant for the conformational change was (4.4 +/- 0.8) X 10(-4) s-1.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Strickland DK,Steiner JP,Feldman SR,Pizzo SVdoi
10.1021/bi00321a061subject
Has Abstractpub_date
1984-12-18 00:00:00pages
6679-85issue
26eissn
0006-2960issn
1520-4995journal_volume
23pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The betagamma-crystallin superfamily consists of evolutionarily related proteins with domain topology similar to lens beta- and gamma-crystallins, formed from duplicated Greek key motifs. Ca(2+) binding was found in a few betagamma-crystallin members earlier, although its prevalence and diversity as inherent molecular...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9017076
更新日期:2009-12-29 00:00:00
abstract::Botulinum neurotoxins (BoNTs) elicit flaccid paralysis through cleavage of SNARE proteins within peripheral neurons. There are seven serotypes of the BoNTs, termed A-G, which differ in the SNARE protein and/or site that is cleaved. BoNTs are single-chain toxins that comprise an N-terminal zinc metalloprotease domain t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi602396x
更新日期:2007-03-20 00:00:00
abstract::Earlier thermodynamic studies of the intermolecular interactions between mature oxytocin and neurophysin, and of the effects of these interactions on neurophysin folding, raised questions about the intramolecular interactions of oxytocin with neurophysin within their common precursor. To address this issue, the disulf...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9912950
更新日期:1999-10-12 00:00:00
abstract::To understand the details of rate limitation of turnover of the photosynthetic reaction center, photooxidation of horse heart cytochrome c by reaction center from Rhodobacter spheroides in detergent dispersion has been examined by intense continuous illumination under a wide variety of conditions of cytochrome concent...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi991563u
更新日期:1999-12-21 00:00:00
abstract::PQQ is an exogenous, tricyclic, quino-cofactor for a number of bacterial dehydrogenases. The final step of PQQ formation is catalyzed by PqqC, a cofactorless oxidase. This study focuses on the activation of molecular oxygen in an enzyme active site without metal or cofactor and has identified a specific oxygen binding...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1015474
更新日期:2011-03-08 00:00:00
abstract::We have used a fluorescent analogue of ATP, mantATP [2'(3')-O-(N-methylanthraniloyl)-adenosine 5'-triphosphate; Hiratsuka T. (1983) Biochim. Biophys. Acta 742, 496-508], and made a detailed kinetic study of the interaction of mantATP and mantADP with S1 and acto-S1. We have shown that these analogues behave like ATP a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00216a017
更新日期:1991-01-15 00:00:00
abstract::An inhibitor of the insulin receptor tyrosine kinase (IRTK), (hydroxy-2-naphthalenyl-methyl) phosphonic acid, was designed and synthesized and was shown to be an inhibitor of the biological effects of insulin in vitro. With a wheat germ purified human placental insulin receptor preparation, this compound inhibited the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00439a053
更新日期:1989-06-27 00:00:00
abstract::The occupancy of the central cavity of the membrane-embedded c ring of the ATP synthase of Escherichia coli was investigated with a photo-cross-linking approach. Single cysteine mutants were created at c subunit positions 4, 8, and 11, which are oriented to the inside of the ring. These cysteines were alkylated with r...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052304+
更新日期:2006-02-14 00:00:00
abstract::The cystic fibrosis transmembrane conductance regulator (CFTR) functions in vivo as a cAMP-activated chloride channel. A member of the ATP-binding cassette superfamily of membrane transporters, CFTR contains two transmembrane domains (TMDs), two nucleotide-binding domains (NBDs), and a regulatory (R) domain. It is pre...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035382a
更新日期:2004-02-03 00:00:00
abstract::Uncontrolled proteolysis due to cathepsin G (cat G) may cause severe pathological disorders. Cat G is inhibited by alpha 1-antichymotrypsin (ACT) and alpha 1-proteinase inhibitor (alpha 1PI), two members of the serpin superfamily of proteins. To see whether these two inhibitors play a physiological proteolysis-prevent...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980223q
更新日期:1998-08-11 00:00:00
abstract::Cyanide inhibits the CO oxidation activity of carbon monoxide dehydrogenase from Clostridium thermoaceticum by binding tightly to the form of the C-cluster yielding the gav = 1.82 signal (the C1.82 form). CN- dissociates and the enzyme reactivates upon addition of CO, CO2 plus dithionite, or CS2 plus dithionite. Dithi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00195a011
更新日期:1994-07-26 00:00:00
abstract::Heparanase is a beta-D-endoglucuronidase that cleaves heparan sulfate (HS) and has been implicated in many important physiological and pathological processes, including tumor cell metastasis, angiogenesis, and leukocyte migration. We report herein the identification of active-site residues of human heparanase. Using P...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002080p
更新日期:2000-12-26 00:00:00
abstract::Continuum methods were used to calculate the electrostatic contributions of charged and polar side chains to the overall stability of a small 41-residue helical protein, the peripheral subunit-binding domain. The results of these calculations suggest several residues that are destabilizing, relative to hydrophobic iso...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992091m
更新日期:2000-02-08 00:00:00
abstract::To determine the active site cysteine residue in aldehyde dehydrogenase, we mutated amino acid residues 49, 162, and 302 of recombinantly expressed rat liver mitochondrial (class 2) aldehyde dehydrogenase. The C49A and C162A mutants were fully active tetrameric enzymes, although the C162A mutant was found to be highly...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00008a025
更新日期:1995-02-28 00:00:00
abstract::The epidermal growth factor receptor is a transmembrane glycoprotein that mediates the cellular responses to epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha). In this study of the human EGF receptor naturally expressed in A431 cells, the glycosylation sites of the full-length, membrane-bo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi027101p
更新日期:2003-05-13 00:00:00
abstract::Manganese superoxide dismutase (MnSOD) cycles between the Mn(II) and Mn(III) states during the catalyzed disproportionation of O(2)(*-), a catalysis that is limited at micromolar levels of superoxide by a peroxide-inhibited complex with the metal. We have investigated the role in catalysis and inhibition of the conser...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011047f
更新日期:2001-10-09 00:00:00
abstract::Hepatic lipase (HL) hydrolysis of phosphatidylcholine (PC) was studied in recombinant high-density lipoprotein particles (r-HDL). r-HDL were made from cholate mixed micelles that contained PC, apo AI, and, in some cases, unesterified cholesterol. r-HDL were characterized using chemical composition, nondenaturing gradi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970356w
更新日期:1997-10-07 00:00:00
abstract::The HIV-1 virulence factor Nef interacts with the macrophage Src-family kinase Hck, resulting in constitutive kinase activation that contributes to viral replication and immune escape. Previous chemical library screens identified the diphenylfuranopyrimdine kinase inhibitor DFP-4AB, which selectively inhibits Nef-depe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00875
更新日期:2015-10-20 00:00:00
abstract::An amplified Okayama-Berg plasmid cDNA library was constructed from total poly(A)+ RNA isolated from the Madin-Darby bovine kidney cell line MDBK. This library was screened with a partial murine calpactin I heavy chain (p36) cDNA clone, the identification of which was based on bovine p36 tryptic peptide sequences gene...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00364a007
更新日期:1986-08-12 00:00:00
abstract::The oxidation of deoxy- and oxyhemerythrin to methemerythrin by Fe(CN)6(3-) has been examined. The first step produces (semi-met)O, a half-oxidized form of hemerythrin which has different spectral and kinetic properties from (semi-met)R which is produced by one-electron reduction of methemerythrin. Further oxidation o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00559a003
更新日期:1980-09-02 00:00:00
abstract::We report differences in the interaction of two structurally similar phenanthroline intercalators, ethidium and propidium, with poly(dA).poly(dT) and poly[d(A-T)] as a function of ionic strength based on titration microcalorimetry, fluorescence titration, and hydrostatic pressure measurements. Both ethidium and propid...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00472a009
更新日期:1990-05-22 00:00:00
abstract::Nuclear magnetic resonance (NMR) (15)N relaxation methods have been used to characterize the backbone dynamics of the N-terminal core domain of the HIV-1 capsid protein (CA(151)). The domain, which has an unusually flat, triangular shape, tumbles in solution at 28 degrees C with an effective rotational correlation tim...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990991x
更新日期:1999-08-10 00:00:00
abstract::The self-association properties of recombinant DNA derived human relaxin, and porcine relaxin isolated from porcine ovaries, have been studied by sedimentation equilibrium analytical ultracentrifugation and circular dichroism (CD). The human relaxin ultracentrifuge data were adequately defined by a monomer-dimer self-...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00245a006
更新日期:1991-08-06 00:00:00
abstract::We previously reported that the microtubule (MT)-binding domain of microtubule-associated protein 4 (MAP4) contains three 18-amino acid imperfect repeats that are homologous to the repeats found in the MT-binding domains of the neuronal MAPs, MAP2 and tau [Chapin, S. J., & Bulinski, J. C. (1991) J. Cell Sci. 98, 27-36...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00007a025
更新日期:1995-02-21 00:00:00
abstract::The roles of four of the active center groups with potential acid-base properties in the region of pH optimum of pyruvate decarboxylase from Saccharomyces cerevisiae have been studied with the substitutions Asp28Ala, His114Phe, His115Phe, and Glu477Gln, introduced by site-directed mutagenesis methods. The steady-state...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002855u
更新日期:2001-06-26 00:00:00
abstract::We have characterized the importance of size, sulfation, and anticoagulant activity of heparin in release of basic fibroblast growth factor (bFGF) from the subendothelial extracellular matrix (ECM) and the luminal surface of the vascular endothelium. For this purpose, 125I-bFGF was first incubated with ECM and conflue...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00122a027
更新日期:1992-02-25 00:00:00
abstract::Lysyl-tRNA synthetase is a member of the class II aminoacyl-tRNA synthetases and catalyses the specific aminoacylation of tRNA(Lys). The crystal structure of the constitutive lysyl-tRNA synthetase (LysS) from Escherichia coli has been determined to 2.7 A resolution in the unliganded form and in a complex with the lysi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001487r
更新日期:2000-10-24 00:00:00
abstract::The amino acid composition and sequence of EIIMtl is known [Lee, C. A., & Saier, M. H., Jr. (1983) J. Biol. Chem. 258, 10761-10767]. This information was combined, in the present study, with quantitative amino acid analysis to determine the molar concentration of the enzyme. The stoichiometry of phosphoryl group incor...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00415a020
更新日期:1988-07-26 00:00:00
abstract::Much interest is currently focused on understanding the detailed contribution that particular amino acid residues make in protein structure and function. Although the use of site-directed mutagenesis has greatly contributed to this goal, the approach is limited to the standard repertoire of twenty amino acids. Fluorin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9617973
更新日期:1997-03-18 00:00:00
abstract::A bifunctional molecule was genetically engineered which contained an amino-terminal effector domain that bound immunoglobulin Fc (fragment B of staphylococcal protein A) and a carboxyl-terminal domain that bound digoxin [a single-chain Fv (sFv)]. Effector and sFv binding properties were virtually identical with those...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00487a005
更新日期:1990-09-04 00:00:00