Abstract:
:Protein kinase C-related kinases (PRKs) are serine/threonine kinases that are members of the protein kinase C superfamily and can be activated by binding to members of the Rho family of small G proteins via a Rho binding motif known as an HR1 domain. The PRKs contain three tandem HR1 domains at their N-termini. The structure of the HR1a domain from PRK1 in complex with RhoA [Maesaki, R., et al. (1999) Mol. Cell 4, 793-803] identified two potential contact interfaces between the G protein and the HR1a domain. In this work, we have used an alanine scanning mutagenesis approach to identify whether both contact sites are used when the two proteins interact in solution and also whether HR1b, the second HR1 domain from PRK1, plays a role in binding to RhoA. The mutagenesis identified just one contact site as being relevant for binding of RhoA and HR1a in solution, and the HR1b domain was found not to contribute to RhoA binding. The folded state and thermal stability of the HR1a and HR1b domains were also investigated. HR1b was found to be more thermally stable than HR1a, and it is hypothesized that the differences in the biophysical properties of these two domains govern their interaction with small G proteins.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hutchinson CL,Lowe PN,McLaughlin SH,Mott HR,Owen Ddoi
10.1021/bi200039usubject
Has Abstractpub_date
2011-04-12 00:00:00pages
2860-9issue
14eissn
0006-2960issn
1520-4995journal_volume
50pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::A new series of negatively charged, paramagnetic alkylsulfonate probes was synthesized and can be used to measure both the internal and the external surface potentials of model membrane systems. We tested for discreteness-of-charge effects in lipid membranes by comparing the surface potentials, estimated by use of the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00373a014
更新日期:1986-12-16 00:00:00
abstract::The inhibition of phosphorylase kinase catalytic activity by 0.1 M neutral salts was predicted by the Hofmeister series of anions. The site of action of the salts was determined by the following evidence to be on the phosphorylase kinase molecule directly, rather than on its protein substrate. (1) Nonactivated kinase ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00665a022
更新日期:1976-10-05 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::We describe a reconstitution of light-activated vertebrate photoreceptor GTPase and a purification of the GTP-binding protein (G protein), which is a component of the GTPase and modulates the light-activated phosphodiesterase (PDE) enzyme system. Rod outer segments (ROS) of bull frogs were treated with ethylenediamine...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00528a003
更新日期:1981-12-08 00:00:00
abstract::Deoxycytidine kinase (dCK) uses either ATP or UTP as a phosphoryl donor to catalyze the phosphorylation of nucleoside acceptors. The kinetic properties of human dCK are modulated in vivo by phosphorylation of serine 74. This residue is a part of the insert region and is distant from the active site. Replacing the seri...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2001032
更新日期:2011-04-12 00:00:00
abstract::A methyltetrahydrofolate:corrinoid/iron-sulfur protein methyltransferase (MeTr) from Clostridium thermoaceticum catalyzes the transfer of the N5 methyl group from (6S)-methyltetrahydrofolate (CH3-H4folate) to the cobalt center of a corrinoid/iron-sulfur protein (C/Fe-SP). The methylcobamide product is the first in a s...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00046a013
更新日期:1995-11-21 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::Using heteronuclear NMR spectroscopy, we demonstrate that a 13-residue peptide (MS-QIKRLLSEKKT) from the cytoplasmic tail of CD4 binds to Nef protein. This part of CD4 is critical for downregulation of CD4 by HIV-1 Nef [Aiken et al. (1994) Cell 76, 853-864]. We show that a control peptide without the central dileucine...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9611164
更新日期:1996-08-13 00:00:00
abstract::A major issue for organisms living at extreme temperatures is to preserve both stability and activity of their enzymes. Cold-adapted enzymes generally have a reduced thermal stability, to counteract freezing, and show a lower enthalpy and a more negative entropy of activation compared to mesophilic and thermophilic ho...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801177k
更新日期:2008-09-23 00:00:00
abstract::The cAMP-dependent protein kinase (cAPK) plays an essential role during differentiation and fruit morphogenesis in Dictyostelium discoideum. The presence of an open reading frame on the gene, pkaC (previously named either Dd PK2 or Dd PK3 by different groups), predicts a 73-kDa polypeptide with 54% similarity to the c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00088a003
更新日期:1993-09-21 00:00:00
abstract::Pili are surface-linked virulence factors that play key roles in infection establishment in a variety of pathogenic species. In Gram-positive pathogens, pilus formation requires the action of sortases, dedicated transpeptidases that covalently associate pilus building blocks. In Streptococcus pneumoniae, a major human...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901261y
更新日期:2009-11-10 00:00:00
abstract::A new class of bacterial multisubunit membrane-bound electron-transfer complexes has been identified based on biochemical and bioinformatic data. It contains subunits homologous to the three-subunit molybdopterin oxidoreductases and four additional subunits, two of which are c-type cytochromes. The complex was purifie...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047267l
更新日期:2005-08-02 00:00:00
abstract::The denaturation of the heat shock protein DnaK induced by guanidine hydrochloride (Gdn-HCl) was investigated by circular dichroism, fluorescence, size-exclusion HPLC, and dynamic light scattering. DnaK unfolding takes place in two discrete steps. The midpoint (Cm) of the first transition (0.5 M) was shifted to higher...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00067a021
更新日期:1993-04-27 00:00:00
abstract::It is now well established that guanine nucleotides are allosteric effectors of the vinca alkaloid-induced self-association of tubulin. GDP enhances self-association for vinblastine-, vincristine- and vinorelbine-induced spiral assembly relative to GTP by 0.90 +/- 0.17 kcal/mol [Lobert et al. (1996) Biochemistry 35, 6...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971120v
更新日期:1997-10-21 00:00:00
abstract::Sulfhydryl-reactive derivatives of glucosamine were synthesized as potentially transportable affinity labels of the human erythrocyte hexose carrier. N-Maleoylglycyl derivatives of either 6- or 2-amino-2-deoxy-D-glucopyranose were the most potent inhibitors of 3-O-methylglucose uptake, with concentrations of half-maxi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00430a044
更新日期:1989-02-21 00:00:00
abstract::The pH dependence of the 13C chemical shifts of the side-chain carboxyl carbons of all Asp and Glu residues in the reduced and oxidized states of human thioredoxin and in a mixed disulfide complex of human thioredoxin with a target peptide from the transcription factor NF kappa B has been investigated by multidimensio...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952299h
更新日期:1996-01-09 00:00:00
abstract::The post-polyketide synthase (PKS) biosynthetic tailoring of macrolide antibiotics usually involves one or more oxidation reactions catalyzed by cytochrome P450 monooxygenases. As the specificities of members from this class of enzymes vary significantly among PKS gene clusters, the identification and study of new mac...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981699c
更新日期:1998-10-20 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 fluorescence spectrum of C1 inhibitor (C1-Inh) in aqueous buffer has a maximum at 324 nm which shifts to 358 nm in 6.0 M guanidinium chloride (GdmC1), indicating that fluorescent tryptophans are buried in the native protein. When titrated with GdmC1, the fluorescence intensity, polarization, and emission maximum o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00331a025
更新日期:1985-05-07 00:00:00
abstract::The S2 subsite specificity of the plant protease papain has been altered to resemble that of mammalian cathepsin B by site-directed mutagenesis. On the basis of amino acid sequence alignments for papain and cathepsin B, a double mutant (Val133Ala/Ser205Glu) was produced where Val133 and Ser205 are replaced by Ala and ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00101a003
更新日期:1991-09-17 00:00:00
abstract::The identity of the sites of insulin-stimulated serine phosphorylation in the human insulin receptor was examined by synthesizing peptides that together encompassed all the serine residues of the cytosolic portion of the beta-subunit and testing them as substrates for phosphorylation by a preparation of human insulin ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00029a025
更新日期:1995-07-25 00:00:00
abstract::Our earlier NMR study showed that a two-disulfide variant of hen lysozyme containing intra-alpha-domain disulfide bridges, C6-C127 and C30-C115, is partially folded, with the alpha domain tightly folded to the nativelike conformation and the beta domain flexible or unfolded. With a view that the formation of a third d...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049967w
更新日期:2004-06-01 00:00:00
abstract::In this study, we have investigated the effect of hydrophobic mismatch between the thickness of the membrane and a transmembrane segment of a protein that directly inserts into the membrane bilayer. For this purpose we used mutants of the single-spanning Pf3 coat protein that can spontaneously insert into Escherichia ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0158674
更新日期:2002-04-16 00:00:00
abstract::Enzyme divergence within three species of the genus Photobacterium (P. fischeri, P. leiognathi, and P. phosphoreum) was studied by comparing the catalytic characteristics and quaternary interactions of bacterial luciferases isolated from each species. Each luciferase was composed of two subunits of different molecular...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00563a009
更新日期:1980-10-28 00:00:00
abstract::The selective reaction of SH containing proteins and peptides with NTCB (2-nitro-5-thiocyanobenzoic acid) has been reported (Degani, Y., & Patchornick, A. (1974) Biochemistry 13, 1; Jacobson, G.A., Schaffer, M.H., Stark, G.R., & Vanaman, T.C. (1973) J. Biol. Chem. 248, 6583). With this reagent, cysteinyl peptide bonds...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00618a022
更新日期:1978-12-12 00:00:00
abstract::Putative domain--domain interactions of the monomeric bifunctional enzyme indoleglycerol phosphate synthase:phosphoribosyl anthranilate isomerase from Escherichia coli were probed by separating the domains on the gene level and expressing them as monofunctional proteins. The engineered monofunctional enzymes were foun...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00016a013
更新日期:1995-04-25 00:00:00
abstract::Thermodynamic equilibrium transition models in DSC are only applicable to reversible processes, but reversibility of the thermal transitions of proteins is comparatively rare because of intermolecular aggregation of denatured proteins and the degradation that occurs at high temperatures. The cupredoxin azurin from Pse...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0157621
更新日期:2002-01-22 00:00:00
abstract::Cyclobutane pyrimidine dimers (Pyr < > Pyr) are the major DNA photoproducts induced by the UV component of solar radiation. Photoreactivating enzyme (DNA photolyase) repairs DNA by utilizing the energy of visible light to break the cyclobutane ring of the dimer. Photolyases are monomeric proteins of 50-60 kDa with sto...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi00167a001
更新日期:1994-01-11 00:00:00
abstract::We have studied the hydrogen-exchange kinetics of hemoglobin A0 as a function of ligand, CO, saturation. In the noncooperative system, azide binding to methemoglobin, the alterations in exchange kinetics are proportional to the average degree of ligation. However, in the case of CO binding to deoxyhemoglobin the chang...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00297a015
更新日期:1984-01-17 00:00:00
abstract::Ultrafast laser spectroscopy techniques are used to measure the low-frequency vibrational coherence spectra and nitric oxide rebinding kinetics of Caldariomyces fumago chloroperoxidase (CPO). Comparisons of the CPO coherence spectra with those of other heme species are made to gauge the protein-specific nature of the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7025485
更新日期:2008-05-06 00:00:00