Abstract:
:The regulatory (R) subunit of cAMP-dependent protein kinase (cAPK) is a multidomain protein with two tandem cAMP-binding domains, A and B. The importance of cAMP binding on the stability of the R subunit was probed by intrinsic fluorescence and circular dichroism (CD) in the presence and absence of urea. Several mutants were characterized. The site-specific mutants R(R209K) and R(R333K) had defects in cAMP-binding sites A and B, respectively. R(M329W) had an additional tryptophan in domain B. Delta(260-379)R lacked Trp260 and domain B. The most destabilizing mutation was R209K. Both CD and fluorescence experiments carried out in the presence of urea showed a decrease in cooperativity of the unfolding, which also occurred at lower urea concentrations. Unlike native R, R(R209K) was not stabilized by excess cAMP. Additionally, CD revealed significant alterations in the secondary structure of the R209K mutant. Therefore, Arg209 is important not only as a contact site for cAMP binding but also for the intrinsic structural stability of the full-length protein. Introducing the comparable mutation into domain B, R333K, had a smaller effect on the integrity and stability of domain A. Unfolding was still cooperative; the protein was stabilized by excess cAMP, but the unfolding curve was biphasic. The R(M329W) mutant behaved functionally like the native protein. The Delta(260-379)R deletion mutant was not significantly different from wild-type RIalpha in its stability. Consequently, domain B and the interaction between Trp260 and cAMP bound to site A are not critical requirements for the structural stability of the cAPK regulatory subunit.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Cànaves JM,Leon DA,Taylor SSdoi
10.1021/bi001563qsubject
Has Abstractpub_date
2000-12-12 00:00:00pages
15022-31issue
49eissn
0006-2960issn
1520-4995pii
bi001563qjournal_volume
39pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全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::Dog serum and colostral immunoglobulin A (IgA) and free secretory component from colostrum were isolated using affinity chromatography. Both serum and colostral IgA showed similar susceptibility to reduction with dithiothreitol, but only colostral IgA released the additional subunit, bound secretory component. This re...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00684a010
更新日期:1975-07-01 00:00:00
abstract::Defects in the MMACHC gene represent the most common disorder of cobalamin (Cbl) metabolism, affecting synthesis of the enzyme cofactors adenosyl-Cbl and methyl-Cbl. The encoded MMACHC protein binds intracellular Cbl derivatives with different upper axial ligands and exhibits flavin mononucleotide (FMN)-dependent decy...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300150y
更新日期:2012-06-26 00:00:00
abstract::His12 and His119 are critical for catalysis of RNA cleavage by ribonuclease A (RNase A). Substitution of either residue with an alanine decreases the value of k(cat)/K(M) by more than 10(4)-fold. His12 and His119 are proximal to the scissile phosphoryl group of an RNA substrate in enzyme-substrate complexes. Here, the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0100182
更新日期:2001-04-24 00:00:00
abstract::The flavoenzyme tryptophan 2-monooxygenase catalyzes the oxidation of tryptophan to indoleacetamide, carbon dioxide, and water. The enzyme is a homologue of l-amino acid oxidase. In the structure of l-amino acid oxidase complexed with aminobenzoate, Tyr372 hydrogen bonds with the carboxylate of the inhibitor in the ac...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035300i
更新日期:2003-12-02 00:00:00
abstract::Targeting damage to specific sites in the genome represents an attractive approach to manipulating gene function in mammalian cells. To test the applicability of triple-helix formation as a means for achieving precisely timed site-specific damage within a mammalian gene, a triplex-forming oligodeoxyribonucleotide (TFO...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960881f
更新日期:1996-08-20 00:00:00
abstract::The antiviral and antiproliferative activities of type I interferons (IFNs) are mediated by a common receptor, and its second subunit (IFNAR2) exhibits nanomolar affinity to both IFNalpha and IFNbeta subtypes. We have previously determined the structure of the IFN-binding extracellular domain of IFNAR2 (IFNAR2-EC) usi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049606g
更新日期:2004-08-10 00:00:00
abstract::We report cloning, expression in Escherichia coli, and purification of cytochrome P450 from a deep-sea bacterium Photobacterium profundum strain SS9 (P450-SS9). The enzyme, which is predominately high spin (86%) in the absence of any added ligand, binds fatty acids and their derivatives and exhibits the highest affini...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi101466y
更新日期:2010-12-21 00:00:00
abstract::The membrane-active, cationic, β-hairpin peptide, arenicin, isolated from marine polychaeta Arenicola marina exhibits a broad spectrum of antimicrobial activity. The peptide in aqueous solution adopts the significantly twisted β-hairpin conformation without pronounced amphipathicity. To assess the mechanism of arenici...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200746t
更新日期:2011-07-19 00:00:00
abstract::Using our hemoglobin expression system in Escherichia coli, we have constructed three recombinant hemoglobins (rHbs) with amino acid substitutions located in the alpha(1)beta(1) and alpha(1)beta(2) subunit interfaces and in the distal heme pocket of the alpha-chain: rHb(alphaV96W, betaN108K), rHb(alphaL29F, alphaV96W,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi991271t
更新日期:1999-10-05 00:00:00
abstract::ATP-dependent Mg2+ accumulation in isolated mitochondria occurs predominantly in the matrix and inner membrane compartments. In mitochondria contaminated with lysosomes, the time course and magnitude of ATP-dependent Mg2+ accumulation are influenced by various cytoplasmic substances, besides substrates of the citric a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00656a013
更新日期:1976-06-01 00:00:00
abstract::The beta subunits of the 143-kDa alpha2beta2 tetrameric enzyme tryptophan synthase have been labeled by L-[ring-4-19F]phenylalanine and L-[phenol-4-13C]tyrosine in an effort to monitor the positions of these residues on ligand binding. Of the 13 phenylalanine and 11 tyrosine residues in the beta subunit, only three pa...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9518297
更新日期:1996-03-12 00:00:00
abstract::Papain-solubilized human class II (HLA-DR) antigens have been purified from cadaveric spleens by ion-exchange chromatography, gel chromatography, and immunosorbent purification. The isolated papain-solubilized antigens comprised two subunits with apparent molecular weights of 23 000 and 30 000, respectively. The circu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00264a036
更新日期:1982-10-12 00:00:00
abstract::Fluoroaluminate and fluoroberyllate are potent inhibitors of the ATPase activity of myosin. Inhibition requires the presence of ADP, and much evidence has accumulated to suggest that the tetrahedral fluoroaluminate and fluoroberyllate ions act as phosphate analogues, binding with high affinity at the active site in th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00090a022
更新日期:1993-10-05 00:00:00
abstract::Aspartate transcarbamoylase from Escherichia coli is a dodecameric enzyme consisting of two trimeric catalytic subunits and three dimeric regulatory subunits. The X-ray structure of this enzyme indicates that the side chains of His-41, Asp-100, and Asp-90 from one catalytic chain form interactions with the side chains...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00089a034
更新日期:1993-09-28 00:00:00
abstract::The mechanism of activation of cGMP phosphodiesterase by the GTP-binding protein in the disc membrane of retinal rods has been investigated by measuring the light-induced phosphodiesterase activity in reconstituted systems where the concentration of either the GTP-binding protein or the phosphodiesterase is varied. Th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00444a040
更新日期:1989-09-05 00:00:00
abstract::This study presents an approach to identifying surface residues on membrane proteins that are exposed toward the membrane-aqueous interface. The method employs a lipid Ni(II) chelate that localizes the metal ion to a region near the membrane-aqueous interface. Lateral diffusion of the lipid chelate results in Heisenbe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi015828s
更新日期:2002-01-29 00:00:00
abstract::The [2Fe-2S] cluster of the Rieske iron-sulfur protein is held between two loops of the protein that are connected by a disulfide bridge. We have replaced the two cysteines that form the disulfide bridge in the Rieske protein of Saccharomyces cerevisiae with tyrosine and leucine, and tyrosine and valine, to evaluate t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035344r
更新日期:2003-11-25 00:00:00
abstract::Thrombospondin-1 (TSP1) binding to calreticulin (CRT) on the cell surface signals focal adhesion disassembly, leading to the intermediate adhesive phenotype, cell migration, anoikis resistance, and collagen stimulation. Residues Lys 24 and 32 in TSP1 and amino acids 24-26 and 32-34 in CRT have been shown through bioch...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi101639y
更新日期:2011-02-01 00:00:00
abstract::A considerable body of data supports the model that the infectious agent (called a prion) which causes the transmissible spongiform encephalopathies is a replicating polypeptide devoid of nucleic acid. Prions are believed to propagate by changing the conformation of the normal cellular prion protein (PrPc) into an inf...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982328z
更新日期:1999-03-16 00:00:00
abstract::Coenzyme A (CoASH) is the major low-molecular weight thiol in Staphylococcus aureus and a number of other bacteria; the crystal structure of the S. aureus coenzyme A-disulfide reductase (CoADR), which maintains the reduced intracellular state of CoASH, has recently been reported [Mallett, T.C., Wallen, J.R., Karplus, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062299p
更新日期:2007-03-20 00:00:00
abstract::The channel-forming protein OmpF porin from Escherichia coli spans the bacterial outer membrane. Each of the three monomers comprises a hollow, 16-stranded beta-barrel. These are associated to homotrimers which are unusually stable, due mostly to hydrophobic interactions between the beta-barrels. In addition, a loop, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981215c
更新日期:1998-11-10 00:00:00
abstract::SHAPE-MaP is unique among RNA structure probing strategies in that it both measures flexibility at single-nucleotide resolution and quantifies the uncertainties in these measurements. We report a straightforward analytical framework that incorporates these uncertainties to allow detection of RNA structural differences...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00977
更新日期:2015-11-24 00:00:00
abstract::The (beta alpha)8 (or "TIM")-barrel protein phosphoribosylanthranilate isomerase from Saccharomyces cerevisiae was cleaved between the sixth and seventh beta alpha module to test the capacity of the resulting fragments to adopt native format autonomously. The fragments, which were expressed from separate coding sequen...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00129a010
更新日期:1992-04-14 00:00:00
abstract::The acetoacetate decarboxylase-like superfamily (ADCSF) is a little-explored group of enzymes that may contain new biocatalysts. The low level of sequence identity (∼20%) between many ADCSF enzymes and the confirmed acetoacetate decarboxylases led us to investigate the degree of diversity in the reaction and substrate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.9b00652
更新日期:2019-10-08 00:00:00
abstract::In reticulocytes, a single ribosomal protein, S13, has been shown to be phosphorylated by the cAMP-regulated protein kinases. The 40S ribosomal subunits were phosphorylated in vitro with [gamma-32P]ATP to facilitate the identification of S13 during the two-step purification procedure. Total ribosomal protein from the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00604a003
更新日期:1978-05-30 00:00:00
abstract::The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. The sequence was solved primarily by analysis of peptides derived by cleavage at either ly...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00371a048
更新日期:1986-11-18 00:00:00
abstract::GlgE (EC 2.4.99.16) is an α-maltose 1-phosphate:(1→4)-α-d-glucan 4-α-d-maltosyltransferase of the CAZy glycoside hydrolase 13_3 family. It is the defining enzyme of a bacterial α-glucan biosynthetic pathway and is a genetically validated anti-tuberculosis target. It catalyzes the α-retaining transfer of maltosyl units...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi500183c
更新日期:2014-04-22 00:00:00
abstract::Arginine vasopressin (AVP) is a nonapeptide long known as an endocrine and paracrine regulator of important systemic functions, namely, vasoconstriction, gluconeogenesis, corticosteroidogenesis, and excretion of water and urea. Here we report, for the first time, that AVP specifically inhibits expression of the cyclin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026807g
更新日期:2003-02-25 00:00:00
abstract::X-ray studies of the proofreading 3',5'-exonuclease site of the large (Klenow) fragment of DNA polymerase I have detected a binuclear metal complex consisting of a pentacoordinate metal (site A) which shares a ligand, Asp-355, with an octahedral metal (site B) [Freemont, P. S., Friedman, J. M., Beese, L. S., Sanderson...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00110a012
更新日期:1991-11-19 00:00:00