Studies on transformation of Escherichia coli with plasmids.
AuthorsHanahan D,
JournalJ Mol Biol
PubMed ID6345791
Factors that affect the probability of genetic transformation of Escherichia coli by plasmids have been evaluated. A set of conditions is described under which about one in every 400 plasmid molecules produces a transformed cell. These conditions include cell growth in medium containing elevated levels of Mg2+, and incubation of ... More
DNA sequence variation in the promoter region of the VEGF gene impacts VEGF gene expression and maximal oxygen consumption.
AuthorsPrior SJ, Hagberg JM, Paton CM, Douglass LW, Brown MD, McLenithan JC, Roth SM,
JournalAm J Physiol Heart Circ Physiol
PubMed ID16339827
'In its role as an endothelial cell proliferation and migration factor, vascular endothelial growth factor (VEGF) can affect peripheral circulation, and therefore impact maximal oxygen consumption (Vo2max). Because of the role of VEGF, and because variation in the VEGF gene has the ability to alter VEGF gene expression and VEGF ... More
A role for mitochondrial Bak in apoptotic response to anticancer drugs.
Authors Wang G Q; Gastman B R; Wieckowski E; Goldstein L A; Gambotto A; Kim T H; Fang B; Rabinovitz A; Yin X M; Rabinowich H;
JournalJ Biol Chem
PubMed ID11447222
'In the present study a clonal Jurkat cell line deficient in expression of Bak was used to analyze the role of Bak in cytochrome c release from mitochondria. The Bak-deficient T leukemic cells were resistant to apoptosis induced by UV, staurosporin, VP-16, bleomycin, or cisplatin. In contrast to wild type ... More
Comparison of kinetic properties between two mammalian ras p21 GDP/GTP exchange proteins, ras guanine nucleotide-releasing factor and smg GDP dissociation stimulation.
'The mammalian counterpart of the yeast ras p21 GDP/GTP exchange protein CDC25, ras GRF, was expressed in Escherichia coli and purified, and its kinetic properties were compared with those of another mammalian ras p21 GDP/GTP exchange protein, smg GDS. ras GRF was active on Ki- and Ha- ras p21s but ... More
Residues in the first extracellular loop of a G protein-coupled receptor play a role in signal transduction.
AuthorsAkal-Strader A, Khare S, Xu D, Naider F, Becker JM.
JournalJ Biol Chem
PubMed ID12058045
'The Saccharomyces cerevisiae pheromone, alpha-factor (WHWLQLKPGQPMY), and Ste2p, its G protein-coupled receptor, were used as a model system to study ligand-receptor interaction. Cys-scanning mutagenesis on each residue of EL1, the first extracellular loop of Ste2p, was used to generate a library of 36 mutants with a single Cys residue substitution. ... More
The Domain Organization and Properties of Individual Domains of DNA Topoisomerase V, a Type 1B Topoisomerase with DNA Repair Activities.
Authors Belova Galina I; Prasad Rajendra; Nazimov Igor V; Wilson Samuel H; Slesarev Alexei I;
JournalJ Biol Chem
PubMed ID11733530
'Topoisomerase V (Topo V) is a type IB (eukaryotic-like) DNA topoisomerase. It was discovered in the hyperthermophilic prokaryote Methanopyrus kandleri and is the only topoisomerase with associated apurinic/apyrimidinic (AP) site-processing activities. The structure of Topo V in the free and DNA-bound states was probed by limited proteolysis at 37 degrees ... More
Site-directed mutagenesis of large (13-kb) plasmids in a single-PCR procedure
AuthorsWang J, Wilkinson MF
JournalBiotechniques
PubMed ID11084857
Although there are many methods for site-specific mutagenesis, most are only applicable for short (<7 kb) plasmids. The few methods that have been developed for longer plasmids are complicated procedures that involve multiple PCR steps or complx recombination and ligation reactions. Here, we provide a simple single-PCR ... More
NSF ATPase and alpha-/beta-SNAPs Disassemble the AMPA Receptor-PICK1 Complex.
Authors Hanley Jonathan G; Khatri Latika; Hanson Phyllis I; Ziff Edward B;
JournalNeuron
PubMed ID11931741
AMPA receptor (AMPAR) trafficking is crucial for synaptic plasticity that may be important for learning and memory. NSF and PICK1 bind the AMPAR GluR2 subunit and are involved in trafficking of AMPARs. Here, we show that GluR2, PICK1, NSF, and alpha-/beta-SNAPs form a complex in the presence of ATPgammaS. Similar ... More
Two distinct pseudomonas effector proteins interact with the pto kinase and activate plant immunity.
Authors Kim Young Jin; Lin Nai Chun; Martin Gregory B;
JournalCell
PubMed ID12062102
The Pto serine/threonine kinase of tomato confers resistance to speck disease by recognizing strains of Pseudomonas syringae that express the protein AvrPto. Pto and AvrPto physically interact, and this interaction is required for activation of host resistance. We identified a second Pseudomonas protein, AvrPtoB, that interacts specifically with Pto and ... More
Requirement for either a host- or pectin-induced pectate lyase for infection of pisum sativum by nectriahematococca.
AuthorsRogers LM, Kim YK, Guo W, Gonzalez-Candelas L, Li D, Kolattukudy PE
JournalProc Natl Acad Sci U S A
PubMed ID10931947
Fungal pathogens usually have multiple genes that encode extracellular hydrolytic enzymes that may degrade the physical barriers in their hosts during the invasion process. Nectria hematococca, a plant pathogen, has two inducible pectate lyase (PL) genes (pel) encoding PL that can help degrade the carbohydrate barrier in the host. pelA ... More
Sites important for Na+ and substrate binding in the Na+/proline transporter of Escherichia coli, a member of the Na+/solute symporter family.
To elucidate the functional importance of transmembrane domain II in the Na(+)/proline transporter (PutP) of Escherichia coli we analyzed the effect of replacing Ser-54 through Gly-58. Substitution of Asp-55 or Met-56 dramatically reduces the apparent affinity for Na(+) and Li(+) in a cation-dependent manner. Conversely, Cys in place of Gly-58 ... More
Identification of amino acid residues critical for biological activity in human interleukin-18.
Authors Kim Soo-Hyun; Azam Tania; Novick Daniela; Yoon Do-Young; Reznikov Leonid L; Bufler Philip; Rubinstein Menachem; Dinarello Charles A;
JournalJ Biol Chem
PubMed ID11790772
Interleukin-18 (IL-18) is a pro-inflammatory cytokine, and IL-18-binding protein (IL-18BP) is a naturally occurring protein that binds IL-18 and neutralizes its biological activities. Computer modeling of human IL-18 identified two charged residues, Glu-42 and Lys-89, which interact with oppositely charged amino acid residues buried in a large hydrophobic pocket of ... More