Abstract:
:Beta-2-microglobulin (beta2m) deposits as amyloid fibrils in the musculoskeletal system of patients undergoing long-term dialysis treatment as a result of kidney failure. Previous work has shown that Cu(II) binding causes beta2m to organize into nativelike dimers and tetramers that precede amyloid formation. Cu(II) is then released from higher-order oligomers before mature Cu(II)-free amyloid fibrils are formed. While some of the Cu(II)-induced structural changes that enable beta2m self-assembly are starting to be revealed, the details of how the Cu(II) binding site evolves from the monomer to the dimers and tetramers are not known. Here, we report results from three mass spectrometry (MS)-based methods that provide insight into the changing Cu-beta2m interactions. We find that monomeric beta2m binds Cu(II) via the N-terminal amine, the amide of Gln2, His31, and Asp59. In the dimer and tetramer, Asp59 is no longer bound to Cu(II), but the other residues still comprise a well-defined albeit weaker binding site that is better able to release Cu(II). Consistent with this is the observation that a fraction of the tetrameric species no longer binds Cu(II) at this weakened binding site, which agrees with a previous report that suggested the tetramer as the first Cu(II)-free oligomer. Our results also provide some insight into structural changes caused by Cu(II) binding that facilitate oligomer formation. Specifically, binding by Asp59 in the monomer requires significant movement of this residue, and we propose that this repositioning is important for establishing a pair of dimer-stabilizing salt bridges between this residue and Lys19. We also find evidence that Cu(II) binding in the N-terminal region of the monomer repels Arg3, which likely allows this residue to form a pair of dimer-stabilizing salt bridges with Glu16. Overall, our measurements suggest that the previously proposed conformational switch caused by Cu(II) binding includes not only a cis-trans isomerization at Pro32 but also the repositioning of residues that are critical for the formation of new electrostatic interactions.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Srikanth R,Mendoza VL,Bridgewater JD,Zhang G,Vachet RWdoi
10.1021/bi901172ysubject
Has Abstractpub_date
2009-10-20 00:00:00pages
9871-81issue
41eissn
0006-2960issn
1520-4995journal_volume
48pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The fatty acid synthase (FAS) of animal tissue is a dimer of two identical subunits, each with a Mr of 260,000. The subunit is a single multifunctional protein having seven catalytic activities and a site for binding of the prosthetic group 4'-phosphopantetheine. The mRNA coding for the subunit has an estimated size o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00420a029
更新日期:1988-10-04 00:00:00
abstract::The interaction of bee venom melittin with dimyristolphosphatidylcholine (DMPC) selectively deuteriated in the choline head group has been studied by deuterium and phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy. The action of residual phospholipase A2 in melittin samples resulted in mixtures of DMPC and i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00392a033
更新日期:1987-09-08 00:00:00
abstract::Electron spin polarized electron paramagentic resonance (ESP EPR) spectra were obtained with deuterated iron-removed photosynthetic bacterial reaction centers (RCs) to specifically investigate the effect of the rate of primary charge separation, metal-site occupancy, and H-subunit content on the observed P865+QA- char...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9630319
更新日期:1997-07-15 00:00:00
abstract::Sulfite oxidase (SO) catalyzes the physiologically critical conversion of sulfite to sulfate. Enzymatic activity is dependent on the presence of the metal molybdenum complexed with a pyranopterin-dithiolene cofactor termed molybdopterin. Comparison of the amino acid sequences of SOs from a variety of sources has ident...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1001954
更新日期:2010-05-11 00:00:00
abstract::Phosphorylation by ATP of E.*Ca2 (sarcoplasmic reticulum vesicles (SRV) with bound 45Ca2+) during 5-10 ms leads to the occlusion of 2 *Ca2+/EPtot [quench by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) alone] in both "empty" (10 microM free Ca2+in) or "loaded" SRV (20-40 mM free Ca2+in)...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00408a041
更新日期:1988-04-19 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::Phototropin (phot) is a blue-light sensor protein that elicits several photo responses in plants. Phototropin has two flavin mononucleotide (FMN)-binding domains, LOV1 and LOV2, in its N-terminal half. The C-terminal half is a blue-light-regulated Ser/Thr kinase. Various functional studies have reported that only LOV2...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701851v
更新日期:2008-01-22 00:00:00
abstract::Rhodanese bound to bacterial ribosomes as peptidyl-tRNA can be folded into an enzymatically active conformation by generating C-terminal extensions of the wild-type enzyme. Rhodanese was synthesized by coupled transcription/translation in a cell-free Escherichia coli system from plasmids containing the coding sequence...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00044a003
更新日期:1995-11-07 00:00:00
abstract::Proteases specific for cleavage after acidic residues have been implicated in several disease states, including epidermolysis, inflammation, and viral processing. A serine protease with specificity toward glutamic acid substrates (Glu-SGP) has been crystallized in the presence of a tetrapeptide ligand and its structur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00094a001
更新日期:1993-11-02 00:00:00
abstract::In clostridial-type ferredoxins, each of the two [4Fe-4S]2+/+ clusters receives three of its four ligands from a CysXXCysXXCys motif. Azotobacter vinelandii ferredoxin I (AvFdI) is a seven-iron ferredoxin that contains one [4Fe-4S]2+/+ cluster and one [3Fe-4S]+/0 cluster. During the evolution of the 7Fe azotobacter-ty...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9810499
更新日期:1998-09-15 00:00:00
abstract::The doublet at 850 and 830 cm-1 in the Raman spectra of proteins containing tyrosyl residues has been examined as to its origin and the relation of its components to the environment of the phenyl ring, the state of the phenolic hydroxyl group, and the conformation of the amino acid backbone. Raman spectral studies on ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00693a014
更新日期:1975-11-04 00:00:00
abstract::In refolding of iso-2 cytochrome c from Saccharomyces cerevisiae, there are two slow folding reactions, tau 1a and tau 1b. The slower of the slow reactions, tau 1a = 100-200 s, is observed only by absorbance changes, while tau 1b (10-20-fold faster) is detected by fluorescence changes. The temperature dependence of th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00348a024
更新日期:1985-12-31 00:00:00
abstract::Saccharomyces cerevisiae tRNA and 5S RNA, Escherichia coli 5S RNA, and wheat germ 5S RNA have each been specifically spin-labeled at the 3'-terminal ribose to give morpholino-spin-labeled (MSL) RNAs. Enzymatic hydrolysis with pancreatic RNase, followed by anion-exchange chromatography, confirms the site of attachment ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00268a042
更新日期:1982-12-07 00:00:00
abstract::Thirty single cysteine substitution mutants of T4 lysozyme have been prepared and spin-labeled with a sulfhydryl-specific nitroxide reagent in order to systematically investigate the relationship between nitroxide side-chain mobility and protein structure. The perturbation caused by replacement of a native residue wit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960482k
更新日期:1996-06-18 00:00:00
abstract::Acyl carrier protein (ACP) is the central player in fatty acid (FA) biosynthesis. It covalently binds all FA intermediates and presents them to the enzymes needed for elongation. Bacterial ACP must interact with a large number of proteins, which raises the question of how different acyl-ACPs are recognized and disting...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901713r
更新日期:2010-04-06 00:00:00
abstract::A photoaffinity probe for corticotropin (ACTH) receptors was prepared by selective modification of the single tryptophan residue in ACTH. A new photoreactive agent, 2,4-dinitro-5-azidophenylsulfenyl chloride, was synthesized and used for introducing the photoreactive group into ACTH. 2,4-Dinitro-5-azidophenylsulfenyl-...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00555a027
更新日期:1980-07-08 00:00:00
abstract::Many naturally occurring RNA structures contain single mismatches. However, the algorithms currently used to predict RNA structure from sequence rely on a minimal set of data for single mismatches, most of which occur rather infrequently in nature. As a result, several approximations and assumptions are used to predic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701311c
更新日期:2007-11-20 00:00:00
abstract::Transient contacts between denatured polypeptide chains are likely to play an important part in the initial stages of protein aggregation and fibrillation. To analyze the nature of such contacts, we have carried out a protein engineering study of the 102-residue protein U1A, which aggregates transiently in the wild-ty...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi048509k
更新日期:2004-10-19 00:00:00
abstract::Investigation of the charge distribution for all known members of the PP-fold family of peptides reveals a common pattern characterized by a cluster of negative charges in the beta-turn region and a cluster of positive charges in the receptor-binding region of the peptide. Detailed analysis of the electrostatic proper...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00063a005
更新日期:1993-03-30 00:00:00
abstract::The phosphatidylcholine-preferring phospholipase C from Bacillus cereus (PLCBc) is a 28.5 kDa enzyme with three zinc ions in its active site. Although much is known about the roles that various PLCBc active site amino acids play in binding and catalysis, there is little information about the rate-determining step of t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9821216
更新日期:1999-04-06 00:00:00
abstract::Taq DNA polymerase, one of the first thermostable DNA polymerases to be discovered, has been typecast as a DNA-dependent DNA polymerase commonly employed for PCR. However, Taq polymerase belongs to the same DNA polymerase superfamily as the Molony murine leukemia virus reverse transcriptase and has in the past been sh...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00778
更新日期:2020-12-15 00:00:00
abstract::Binding of the nonhydrolyzable ATP analogue 5'-adenylyl-beta, gamma-imidodiphosphate (ADPNP) to Escherichia coli DNA gyrase can lead to a limited noncatalytic supercoiling of DNA. Here we examine the efficiency of coupling between ADPNP binding and the change in linking number either of positively or negatively superc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952433y
更新日期:1996-02-06 00:00:00
abstract::We have studied the Cu(II), Co(II), and Fe(III) complexes of the antineoplastic drug bleomycin by using electron spin--echo envelope spectroscopy. For all three complexes, nitrogen coordination of the metal ions is demonstrated. For the Cu(II)-- and Co(II)--drug complexes, we have been able to identify imidazole as a ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00509a045
更新日期:1981-03-17 00:00:00
abstract::Exposure of mammalian cells (transformed mouse fibroblasts or rat hepatoma cells) to S-adenosyl-1,8-diamino-3-thiooctane produced profound changes in the intracellular polyamine content. Putrescine was increased and spermidine was decreased, consistent with the inhibition of spermidine synthase by this compound, which...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00263a036
更新日期:1982-09-28 00:00:00
abstract::Eukaryotic initiation factor 5 (eIF5) plays multiple roles in translation initiation. Its N-terminal domain functions as a GTPase-activator protein (GAP) for GTP bound to eIF2, while its C-terminal region nucleates the interactions between multiple translation factors, including eIF1, which acts to inhibit GTP hydroly...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052387u
更新日期:2006-04-11 00:00:00
abstract::Evidence is presented that indicates tropoelastin is derived from a soluble elastin with a molecular weight of 95000. Tropoelastin and its proposed precursor were isolated from the aortas of copper-deficient chicks. Although it is doubtful that the proposed precursor is an initial product of elastin translation, i.e.,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00585a004
更新日期:1979-09-04 00:00:00
abstract::Coenzyme A (CoA)-transferases catalyze transthioesterification reactions involving acyl-CoA substrates, using an active-site carboxylate to form covalent acyl anhydride and CoA thioester adducts. Mechanistic studies of class I CoA-transferases suggested that acyl-CoA binding energy is used to accelerate rate-limiting ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300957f
更新日期:2012-10-23 00:00:00
abstract::The active site of methionyl-tRNA synthetase (MetRS) possesses two functions: synthetic, which provides Met-tRNA for protein synthesis, and editing, which rejects inadvertently misactivated homocysteine. During editing, the side chain -SH group of homocysteine reacts with its activated carboxyl group forming a cyclic ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960344v
更新日期:1996-06-25 00:00:00
abstract::The specificity of horse liver alcohol dehydrogenase for cyclohexanol and its 3-methyl derivatives was investigated by stopped-flow and initial velocity kinetic studies. The (1S,3S)-3-methylcyclohexanol was 7 times more reactive (V/Km) than cyclohexanol, whereas the (1R,3R)-3-methylcyclohexanol was at least 1000 times...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00409a060
更新日期:1988-05-03 00:00:00
abstract::Antisera were raised in rabbits against the 40-kDa alpha subunit of bovine lung GTP-binding protein, which were identified as the alpha subunit of Gi2 (Gi2 alpha) by the analysis of the partial amino acid sequence. Antibodies were purified with a Gi2 alpha-coupled Sepharose column and then were passed through a Gi1 al...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00437a035
更新日期:1989-05-30 00:00:00