Abstract:
:The two key structural features of alpha 2-macroglobulin (alpha 2M) involved in inhibitory caging of proteases are the thiol ester and the bait region. This paper examines the environment of the hydrolyzed thiol ester in methylamine-treated human alpha 2M and the separation between the bait region and the thiol ester and between the four thiol esters in the tetramer to try to further our understanding of how bait region proteolysis triggers thiol ester cleavage. The sulfhydryl groups of Cys-949, formed upon cleavage of the thiol ester by methylamine, were specifically labeled with the nitroxide spin-labels 3-(2-iodoacetamido)-PROXYL (iodo-I) (PROXYL = 2,2,5,5-tetramethylpyrrolidine-1-oxyl), 3-[2-(2-iodoacetamido)acetamido]-PROXYL (iodo-II), and 4-(2-iodoacetamido)-2,2,6,6-tetramethylpiperidine-1-oxyl (iodo-III). ESR spectra of these alpha 2M derivatives showed that label I is firmly held and label II has limited freedom of rotation consistent with location of the cysteine residue in a narrow cavity. Label III has much greater motional freedom. From the absence of dipole-dipole splittings in the ESR spectra, it is concluded that the four nitroxide groups in the tetramer are more than 20 A apart for both label I and label II. Label I broadens 1H NMR signals from one phenylalanyl, one tyrosyl, and four histidyl residues in the bait region. Separations of 11-17 A are estimated between the nitroxide of label I and these residues. Label II is further away and only broadens resonances from one of the histidines.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Gettins P,Beth AH,Cunningham LWdoi
10.1021/bi00408a036subject
Has Abstractpub_date
1988-04-19 00:00:00pages
2905-11issue
8eissn
0006-2960issn
1520-4995journal_volume
27pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Site-specific mutagenesis has been used to create two mutant versions of aspartate transcarbamoylase. Arg-167 and Gln-231, both previously identified as interacting with the portion of the bisubstrate analogue N-(phosphonoacetyl)-L-aspartate (PALA) that corresponds to aspartate [Krause, K. L., Voltz, K. W., & Lipscomb...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00468a003
更新日期:1990-04-24 00:00:00
abstract::To shed more light on the molecular requirements for recognition of thyroid response elements (TREs) by thyroid receptors (TRs), we compared the specific aspects of DNA TRE recognition by different TR constructs. Using fluorescence anisotropy, we performed a detailed and hierarchical study of TR-TRE binding. This was ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901282s
更新日期:2010-02-09 00:00:00
abstract::Bovine seminal ribonuclease (BS-RNase), the only dimeric protein among the pancreatic-like ribonucleases, is endowed with special structural features and with biological functions beyond enzymatic activity. In solution, the protein exists as an equilibrium mixture of two forms, with or without exchange (or swapping) o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0342517
更新日期:2003-07-29 00:00:00
abstract::The 2.15 A resolution crystal structure of arginase from Plasmodium falciparum, the parasite that causes cerebral malaria, is reported in complex with the boronic acid inhibitor 2(S)-amino-6-boronohexanoic acid (ABH) (K(d) = 11 microM). This is the first crystal structure of a parasitic arginase. Various protein const...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100390z
更新日期:2010-07-06 00:00:00
abstract::A critical event in Alzheimer's disease is the transition of Abeta peptides from their soluble forms into disease-associated beta-sheet-rich conformers. Structural analysis of a complete D-amino acid replacement set of Abeta(1-42) enabled us to localize in the full-length 42-mer peptide the region responsible for the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002005e
更新日期:2001-05-08 00:00:00
abstract::The Met16Phe mutant of the type 1 copper protein pseudoazurin (PACu), in which a phenyl ring is introduced close to the imidazole moiety of the His81 ligand, has been characterized. NMR studies indicate that the introduced phenyl ring is parallel to the imidazole group of His81. The mutation has a subtle effect on the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi030030p
更新日期:2003-06-10 00:00:00
abstract::New homo- and heterobifunctional cross-linking reagents have been synthesized. These reagents are based on ortho ester, acetal, and ketal functionalities that undergo acid-catalyzed dissociation but are base stable. The protein-reactive group in all the homobifunctional reagents is a maleimide group; the heterobifunct...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00432a023
更新日期:1989-03-21 00:00:00
abstract::We report the genetically encoded chemical decaging strategy for protein activation in living bacterial cells. In contrast to the metabolically labile photocaging groups inside Escherichia coli, our chemical decaging strategy that relies on the inverse electron-demand Diels-Alder (iDA) reaction is compatible with the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.7b01017
更新日期:2018-01-30 00:00:00
abstract::G protein alpha subunits mediate activation of signaling pathways through G protein-coupled receptors (GPCR) by virtue of GTP-dependent conformational rearrangements. It is known that regions of disorder in crystal structures can be indicative of conformational flexibility within a molecule, and there are several such...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0255726
更新日期:2002-08-06 00:00:00
abstract::The initial hydrolysis of CaATP by chloroplast coupling factor 1 was studied with the quenched-flow method. The time course of hydrolysis can be described as a first-order conversion of the enzyme to an active form followed by steady-state formation of product. The rat constant for the first-order process is independe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00382a035
更新日期:1987-04-21 00:00:00
abstract::Traditional antibody generation, using either phage display or animal immunization, relies on purified antigens. Many membrane proteins, such as G protein-coupled receptors, solute carriers, or ion channels, are important drug targets but very challenging for the formation of antibodies due to the difficulty of protei...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00730
更新日期:2020-11-10 00:00:00
abstract::Factor XIIIa (a2') is a homodimeric transglutaminase that is formed via limited alpha-thrombin-catalyzed proteolysis of the platelet (a2) or plasma (a2b2) factor XIII zymogen in a reaction that results in proteolytic removal of a 37-aminoacyl residue peptide from the N-terminus of the a chains and exposure of the acti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00117a017
更新日期:1992-01-21 00:00:00
abstract::31P NMR measurements were made (at 121.5 MHz and 5 degrees C) on enzyme-bound substrate complexes of 3-phosphoglycerate kinase in order to address three questions pertaining to (i) the integrity of the enzyme-substrate complexes with Mg(II) in the presence of sulfate concentrations typical of those used for crystalliz...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00415a027
更新日期:1988-07-26 00:00:00
abstract::The interaction of fluorescent ATP analog 2'(3')-O-[N-[2-[3-(5-fluoresceinyl)thioureido]-ethyl]carbamoyl]adenosine 5'-triphosphate (FEDA-ATP) with rabbit skeletal myosin subfragment 1 (S1) and acto-S1 was studied. This and related ATP analogs are potentially useful for determination of the ATPase activity of single my...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi951824+
更新日期:1996-02-20 00:00:00
abstract::A cDNA designated hIIB1, representing the entire coding sequence of a P450 in the IIB gene family, was isolated from a human liver lambda gt11 library by using the rat IIB1 cDNA as a probe. The hIIB1 protein, deduced from the cDNA sequence, contained 491 amino acids, had a calculated molecular weight of 56,286, and di...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00444a029
更新日期:1989-09-05 00:00:00
abstract::Intensive study has been devoted to understanding the kinetic and structural bases underlying the exceptionally high fidelity (low error frequencies) of the typical DNA polymerase. Commonly proposed explanations have included (i) the concept of fidelity check points, in which the correctness of a nascent base pair mat...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi026021i
更新日期:2002-08-27 00:00:00
abstract::Binding mechanisms of ADPR-transferase to restricted double-stranded DNA fragments of SV40 and pBR322 DNA were determined by nuclease protection techniques. Top and bottom strands of double-stranded DNA were identified by specific labeling with 32P. Protection against specific exonucleases identified binding of ADPR-t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00439a050
更新日期:1989-06-27 00:00:00
abstract::The mobility of vesicular stomatitis virus (VSV) G protein on the surface of infected BHK cells was studied by using the technique of fluorescence photobleaching recovery. The fraction of surface G protein that was mobile in that time scale of the measurement (minutes) was at least 75%, a relatively high value among c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00508a047
更新日期:1981-03-03 00:00:00
abstract::The initiation of Escherichia coli chromosomal replication by DnaA protein is strongly influenced by the tight binding of the nucleotides ATP and ADP. Anionic phospholipids in a fluid bilayer promote the conversion of inactive ADP-DnaA protein to replicatively active ATP-DnaA protein in vitro, and thus likely play a k...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982733q
更新日期:1999-05-11 00:00:00
abstract::The locations of RNA-RNA cross-links in 16S rRNA were determined after in vivo incorporation of 4-thiouridine (s(4)U) into RNA in a strain of Escherichia coli deficient in pyrimidine synthesis and irradiation at >320 nm. This was done as an effort to find RNA cross-links different from UVB-induced cross-links that wou...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049702h
更新日期:2004-07-20 00:00:00
abstract::The Y70F mutant of aspartate aminotransferase has reduced affinity for coenzymes compared to the wild type. The equilibrium dissociation constants for pyridoxamine phosphate (PMP) holoenzymes, KPMPdiss, were determined from the association and dissociation rate constants to be 1.3 nM and 30 nM for the wild type and mu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00244a014
更新日期:1991-07-30 00:00:00
abstract::1,4-Dihydroxy-2-naphthoyl-coenzyme A (DHNA-CoA) synthase, or MenB, catalyzes an intramolecular Claisen condensation involving two oxyanion intermediates in the biosynthetic pathway of menaquinone, an essential respiration electron transporter in many microorganisms. Here we report the finding that the DHNA-CoA product...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200376x
更新日期:2011-07-05 00:00:00
abstract::1-Bromovinyl (I), Z-2-bromovinyl (II), 1,2-dibromoethyl (III), and a series of 4-(halomethyl)-2-nitrophenyl (IVa-c) diethyl phosphate esters were examined as substrates and mechanism-based inhibitors for the bacterial phosphotriesterase. All of these compounds were found to act as substrates for the enzyme. Inhibitor ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970680r
更新日期:1997-07-22 00:00:00
abstract::High-resolution phosphorus-31 nuclear magnetic resonance (31P NMR) spectra of wild-type and mutant strains of Saccharomyces cerevisiae were observed at a frequency of 145.7 MHz. Levels of various phosphorus metabolites were investigated upon addition of glucose under both aerobic and anaerobic conditions. Three mutant...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00588a006
更新日期:1979-10-16 00:00:00
abstract::The P-glycoprotein multidrug transporter functions as an ATP-driven efflux pump for a large number of structurally unrelated hydrophobic compounds. Substrates are believed to gain access to the transporter after partitioning into the membrane, rather than from the extracellular aqueous phase. The binding of drug subst...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990064q
更新日期:1999-05-25 00:00:00
abstract::alpha-1,3-Fucosyltransferase V (FucT V) catalyzes the transfer of 1-fucose from the donor sugar guanosine 5'-diphospho-beta-1-fucose (GDP-Fuc) to an acceptor sugar. A secondary isotope effect on the fucosyltransfer reaction with guanosine 5'-diphospho-[1-2H]-beta-1-fucose (GDP-[1-2H]-Fuc) as the substrate was observed...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962284z
更新日期:1997-01-28 00:00:00
abstract::The activity of the major isoform of porcine pancreatic phospholipase A2 (PLA2), designated B-PLA2, against micellar substrates is inhibited by heparin. Inhibition is a consequence of binding of the enzyme to heparin, documented by a heparin-induced alteration in the intrinsic fluorescence of B-PLA2 and in the 8-anili...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00101a026
更新日期:1991-09-17 00:00:00
abstract::Room temperature phosphorescence techniques were used to study the structural and dynamic features of the tryptophan residues in bovine alpha-crystallin. Upon excitation at 290 nm, the characteristic signature of tryptophan phosphorescence was observed with an emission maximum at 442 +/- 2 nm. The phosphorescence inte...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00439a027
更新日期:1989-06-27 00:00:00
abstract::Complex I, a key component of the mitochondrial respiratory chain, exhibits diminished activity as a result of cardiac ischemia/reperfusion. Cardiac ischemia/reperfusion is associated with increases in the levels of mitochondrial Ca(2+) and pro-oxidants. In the current in vitro study, we sought evidence for a mechanis...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049803f
更新日期:2004-07-06 00:00:00
abstract::We compared the kinetics and thermodynamics of rhodamine phalloidin binding to actin purified from rabbit skeletal muscle, Acanthamoeba castellanii, and Saccharomyces cerevisiae in 50 mM KCl, 1 mM MgCl2, and pH 7.0 buffer at 22 degrees C. Filaments of S. cerevisiae actin bind rhodamine phalloidin more weakly than Acan...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961047t
更新日期:1996-11-12 00:00:00