Purification and characterization of a ubiquitin carrier protein kinase from HeLa cells.

Abstract:

:Protein ubiquitination plays an important role in ATP-dependent protein turnover, and it also may regulate other cellular events. Covalent attachment of ubiquitin to other proteins is catalyzed by three different enzymes, E1, E2, and E3. We have previously shown that protein ubiquitination can be regulated by phosphorylation. In the present study, we show that 20-kDa E2, an E2 molecular mass isoform, is phosphorylated by a protein kinase from the cytosolic fraction of HeLa cells. This protein kinase was purified by a procedure involving ammonium sulfate precipitation and three column chromatographies (phenyl-Sepharose, Superose gel filtration, and DEAE-Sephacel). Gel-filtration chromatography indicated that the molecular mass of this protein kinase was about 300 kDa. However, SDS/PAGE showed that the purified protein kinase consists of three subunits with molecular masses of 120, 105, and 70 kDa, respectively. The stoichiometry of the phosphorylated 20-kDa E2 isozyme was found to be 0.45 mol of phosphate per mol of protein. The phosphorylation of 20-kDa E2 occurred only at the serine residue. The activity of this protein kinase required the presence of Mg2+; however, the enzyme was inhibited by a high concentration of Mg2+.

authors

Kong SK,Chock PB

doi

10.1073/pnas.91.24.11601

subject

Has Abstract

pub_date

1994-11-22 00:00:00

pages

11601-5

issue

24

eissn

0027-8424

issn

1091-6490

journal_volume

91

pub_type

杂志文章
  • Direct participation of dCMP hydroxymethylase in synthesis of bacteriophage T4 DNA.

    abstract::In order to retain in an in situ system the control mechanisms involved in synthesis of bacteriophage T4 DNA, infected cells were made permeable to nucleotides by plasmolysis with concentrated sucrose. Such preparations use exogenous deoxyribonucleotides to synthesize T4 phage DNA. As has been observed with in vivo st...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.70.8.2196

    authors: Wovcha MG,Tomich PK,Chiu CS,Greenberg GR

    更新日期:1973-08-01 00:00:00

  • Microbial carbohydrate depolymerization by antigen-presenting cells: deamination prior to presentation by the MHCII pathway.

    abstract::After uptake by the endosome of an antigen-presenting cell (APC), exogenous proteins are known to be degraded into peptides by protease digestion. Here, we report the mechanism by which pure carbohydrates can be depolymerized within APC endosomes/lysosomes by nitric oxide (NO)-derived reactive nitrogen species (RNSs) ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0800974105

    authors: Duan J,Avci FY,Kasper DL

    更新日期:2008-04-01 00:00:00

  • Double-core-hole spectroscopy for chemical analysis with an intense X-ray femtosecond laser.

    abstract::Theory predicts that double-core-hole (DCH) spectroscopy can provide a new powerful means of differentiating between similar chemical systems with a sensitivity not hitherto possible. Although DCH ionization on a single site in molecules was recently measured with double- and single-photon absorption, double-core hole...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1111380108

    authors: Berrah N,Fang L,Murphy B,Osipov T,Ueda K,Kukk E,Feifel R,van der Meulen P,Salen P,Schmidt HT,Thomas RD,Larsson M,Richter R,Prince KC,Bozek JD,Bostedt C,Wada S,Piancastelli MN,Tashiro M,Ehara M

    更新日期:2011-10-11 00:00:00

  • Sequence, tissue distribution, and chromosomal localization of mRNA encoding a human glucose transporter-like protein.

    abstract::cDNA clones encoding a glucose transporter-like protein have been isolated from adult human liver and kidney cDNA libraries by cross-hybridization with the human HepG2/erythrocyte glucose transporter cDNA. Analysis of the sequence of this 524-amino acid glucose transporter-like protein indicates that it has 55.5% iden...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.85.15.5434

    authors: Fukumoto H,Seino S,Imura H,Seino Y,Eddy RL,Fukushima Y,Byers MG,Shows TB,Bell GI

    更新日期:1988-08-01 00:00:00

  • Overexpression of normal and mutated forms of HRAS induces orthotopic bladder invasion in a human transitional cell carcinoma.

    abstract::Recent studies have shown that orthotopic (transurethral) transplantation of human bladder cancer cell lines into nude mice permits tumor growth that accurately reflects their clinical malignant status in the original host. Thus, such a system allows a unique opportunity to analyze the genetic events involved in the c...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.22.9047

    authors: Theodorescu D,Cornil I,Fernandez BJ,Kerbel RS

    更新日期:1990-11-01 00:00:00

  • Global regulatory logic for specification of an embryonic cell lineage.

    abstract::Explanation of a process of development must ultimately be couched in the terms of the genomic regulatory code. Specification of an embryonic cell lineage is driven by a network of interactions among genes encoding transcription factors. Here, we present the gene regulatory network (GRN) that directs the specification...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0711220105

    authors: Oliveri P,Tu Q,Davidson EH

    更新日期:2008-04-22 00:00:00

  • Cellular retinol-binding protein allows specific interaction of retinol with the nucleus in vitro.

    abstract::Purified cellular retinol-binding protein (CRBP), a potential mediator of vitamin A action, was found to enable retinol to bind in a specific manner to isolated nuclei from livers of vitamin A deficient rats. Binding was followed after complexing [3H]retinol with CRBP. The binding was specific, saturable, and temperat...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.76.5.2204

    authors: Takase S,Ong DE,Chytil F

    更新日期:1979-05-01 00:00:00

  • lambda Rap protein is a structure-specific endonuclease involved in phage recombination.

    abstract::Bacteriophage lambda encodes a number of genes involved in the recombinational repair of DNA double-strand breaks. The product of one of these genes, rap, has been purified. Truncated Rap proteins that copurify with the full-length form are derived, at least in part, from a rho-dependent transcription terminator locat...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.23.13507

    authors: Sharples GJ,Corbett LM,Graham IR

    更新日期:1998-11-10 00:00:00

  • Evolution of Drosophila sex comb length illustrates the inextricable interplay between selection and variation.

    abstract::In spite of the diversity of possible biological forms observed in nature, a limited range of morphospace is frequently occupied for a given trait. Several mechanisms have been proposed to explain this bias in the distribution of phenotypes including selection, drift, and developmental constraints. Despite extensive w...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1322342111

    authors: Malagón JN,Ahuja A,Sivapatham G,Hung J,Lee J,Muñoz-Gómez SA,Atallah J,Singh RS,Larsen E

    更新日期:2014-09-30 00:00:00

  • Distinct gating mechanism of SOC channel involving STIM-Orai coupling and an intramolecular interaction of Orai in Caenorhabditis elegans.

    abstract::Store-operated calcium entry (SOCE), an important mechanism of Ca2+ signaling in a wide range of cell types, is mediated by stromal interaction molecule (STIM), which senses the depletion of endoplasmic reticulum Ca2+ stores and binds and activates Orai channels in the plasma membrane. This inside-out mechanism of Ca2...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1714986115

    authors: Kim KM,Wijerathne T,Hur JH,Kang UJ,Kim IH,Kweon YC,Lee AR,Jeong SJ,Lee SK,Lee YY,Sim BW,Lee JH,Baig C,Kim SU,Chang KT,Lee KP,Park CY

    更新日期:2018-05-15 00:00:00

  • Ba2LnSbO6 and Sr2LnSbO6 (Ln = Dy, Ho, Gd) double perovskites: lanthanides in the geometrically frustrating fcc lattice.

    abstract::Magnetic ground states in solids often arise as a result of a delicate balance between competing factors. One currently active area of research in magnetic materials involves compounds in which long-range magnetic ordering at low temperatures is frustrated by the geometry of the crystalline lattice, a situation known ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0832394100

    authors: Karunadasa H,Huang Q,Ueland BG,Schiffer P,Cava RJ

    更新日期:2003-07-08 00:00:00

  • Mechanism for pH-dependent gene regulation by amino-terminus-mediated homooligomerization of Bacillus subtilis anti-trp RNA-binding attenuation protein.

    abstract::Anti-TRAP (AT) is a small zinc-binding protein that regulates tryptophan biosynthesis in Bacillus subtilis by binding to tryptophan-bound trp RNA-binding attenuation protein (TRAP), thereby preventing it from binding RNA, and allowing transcription and translation of the trpEDCFBA operon. Crystallographic and sediment...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1004981107

    authors: Sachleben JR,McElroy CA,Gollnick P,Foster MP

    更新日期:2010-08-31 00:00:00

  • Highly accurate two-gene classifier for differentiating gastrointestinal stromal tumors and leiomyosarcomas.

    abstract::Gastrointestinal stromal tumor (GIST) has emerged as a clinically distinct type of sarcoma with frequent overexpression and mutation of the c-Kit oncogene and a favorable response to imatinib mesylate [also known as STI571 (Gleevec)] therapy. However, a significant diagnostic challenge remains in the differentiation o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0611373104

    authors: Price ND,Trent J,El-Naggar AK,Cogdell D,Taylor E,Hunt KK,Pollock RE,Hood L,Shmulevich I,Zhang W

    更新日期:2007-02-27 00:00:00

  • Individual products of the adenovirus 12S and 13S EIa mRNAs stimulate viral EIIa and EIII expression at the transcriptional level.

    abstract::Recombinant plasmids containing mutant or wild-type adenovirus serotype 2 EIa genes that produce the 12S mRNA alone, the 13S mRNA alone, or both mRNAs were cotransfected into HeLa cells with plasmids containing the viral EIIa or EIII transcription units. The amount of RNA produced from the EIIa and EIII promoters was ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.81.14.4381

    authors: Leff T,Elkaim R,Goding CR,Jalinot P,Sassone-Corsi P,Perricaudet M,Kédinger C,Chambon P

    更新日期:1984-07-01 00:00:00

  • Top-down versus bottom-up and the Ruritanian bean bug.

    abstract::In a recent article, Hunter uses the late George Varley and George Gradwell's long-term data on the winter moth (Operophtera brumata) and green tortrix (Tortrix viridana) populations to propose a method of quantifying the relative importance of top-down effects (because of natural enemies) and bottom-up effects (becau...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.18.10661

    authors: Hassell MP,Crawley MJ,Godfray HC,Lawton JH

    更新日期:1998-09-01 00:00:00

  • Room-temperature single-electron junction.

    abstract::The design, realization, and test performances of an electronic junction based on single-electron phenomena that works in the air at room temperature are hereby reported. The element consists of an electrochemically etched sharp tungsten stylus over whose tip a nanometer-size crystal was synthesized. Langmuir-Blodgett...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.20.10556

    authors: Facci P,Erokhin V,Carrara S,Nicolini C

    更新日期:1996-10-01 00:00:00

  • Illusory motion perception in blindsight.

    abstract::Motion detection is typically spared in blindsight, which results from damage to the striate cortex (area V1) of the brain that is sufficient to eliminate conscious visual awareness and severely reduce sensitivity to luminance contrast, especially for high spatial and low temporal frequencies. Here we show that the di...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1005974108

    authors: Azzopardi P,Hock HS

    更新日期:2011-01-11 00:00:00

  • The kuru infectious agent is a unique geographic isolate distinct from Creutzfeldt-Jakob disease and scrapie agents.

    abstract::Human sporadic Creutzfeldt-Jakob disease (sCJD), endemic sheep scrapie, and epidemic bovine spongiform encephalopathy (BSE) are caused by a related group of infectious agents. The new U.K. BSE agent spread to many species, including humans, and clarifying the origin, specificity, virulence, and diversity of these agen...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0905825106

    authors: Manuelidis L,Chakrabarty T,Miyazawa K,Nduom NA,Emmerling K

    更新日期:2009-08-11 00:00:00

  • Vaccines, new opportunities for a new society.

    abstract::Vaccination is the most effective medical intervention ever introduced and, together with clean water and sanitation, it has eliminated a large part of the infectious diseases that once killed millions of people. A recent study concluded that since 1924 in the United States alone, vaccines have prevented 40 million ca...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章,评审

    doi:10.1073/pnas.1402981111

    authors: Rappuoli R,Pizza M,Del Giudice G,De Gregorio E

    更新日期:2014-08-26 00:00:00

  • ChIP-on-chip significance analysis reveals large-scale binding and regulation by human transcription factor oncogenes.

    abstract::ChIP-on-chip has emerged as a powerful tool to dissect the complex network of regulatory interactions between transcription factors and their targets. However, most ChIP-on-chip analysis methods use conservative approaches aimed at minimizing false-positive transcription factor targets. We present a model with improve...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0806445106

    authors: Margolin AA,Palomero T,Sumazin P,Califano A,Ferrando AA,Stolovitzky G

    更新日期:2009-01-06 00:00:00

  • Allosteric regulation of an essential trypanosome polyamine biosynthetic enzyme by a catalytically dead homolog.

    abstract::African sleeping sickness is a fatal disease that is caused by the protozoan parasite Trypanosoma brucei. Polyamine biosynthesis is an essential pathway in the parasite and is a validated drug target for treatment of the disease. S-adenosylmethionine decarboxylase (AdoMetDC) catalyzes a key step in polyamine biosynthe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0701111104

    authors: Willert EK,Fitzpatrick R,Phillips MA

    更新日期:2007-05-15 00:00:00

  • Profiling of tyrosine phosphorylation pathways in human cells using mass spectrometry.

    abstract::The reversible phosphorylation of tyrosine residues is an important mechanism for modulating biological processes such as cellular signaling, differentiation, and growth, and if deregulated, can result in various types of cancer. Therefore, an understanding of these dynamic cellular processes at the molecular level re...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2436191100

    authors: Salomon AR,Ficarro SB,Brill LM,Brinker A,Phung QT,Ericson C,Sauer K,Brock A,Horn DM,Schultz PG,Peters EC

    更新日期:2003-01-21 00:00:00

  • High sensitivity of BRCA1-deficient mammary tumors to the PARP inhibitor AZD2281 alone and in combination with platinum drugs.

    abstract::Whereas target-specific drugs are available for treating ERBB2-overexpressing and hormone receptor-positive breast cancers, no tailored therapy exists for hormone receptor- and ERBB2-negative ("triple-negative") mammary carcinomas. Triple-negative tumors account for 15% of all breast cancers and frequently harbor defe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0806092105

    authors: Rottenberg S,Jaspers JE,Kersbergen A,van der Burg E,Nygren AO,Zander SA,Derksen PW,de Bruin M,Zevenhoven J,Lau A,Boulter R,Cranston A,O'Connor MJ,Martin NM,Borst P,Jonkers J

    更新日期:2008-11-04 00:00:00

  • Immunotoxin SS1P is rapidly removed by proximal tubule cells of kidney, whose damage contributes to albumin loss in urine.

    abstract::Recombinant immunotoxins (RITs) are chimeric proteins composed of an Fv and a protein toxin being developed for cancer treatment. The Fv brings the toxin to the cancer cell, but most of the RITs do not reach the tumor and are removed by other organs. To identify cells responsible for RIT removal, and the pathway by wh...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1919038117

    authors: Liu XF,Wei J,Zhou Q,Molitoris BA,Sandoval R,Kobayashi H,Okada R,Nagaya T,Karim B,Butcher D,Pastan I

    更新日期:2020-03-17 00:00:00

  • Consequences of reproductive barriers for genealogical discordance in the European corn borer.

    abstract::Speciation involves the origin of trait differences that limit or prevent gene exchange and ultimately results in daughter populations that form monophyletic or exclusive genetic groups. However, for recently diverged populations or species between which reproductive isolation is often incomplete, gene genealogies wil...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0502054102

    authors: Dopman EB,Pérez L,Bogdanowicz SM,Harrison RG

    更新日期:2005-10-11 00:00:00

  • Synergistic and promoter-selective activation of transcription by recruitment of transcription factors TFIID and TFIIB.

    abstract::Eukaryotic transcriptional activators may function by stimulating formation of RNA polymerase II preinitiation complexes at the core promoter of genes. In this case, their mode of action will intrinsically depend on how these complexes assemble on promoters in living cells, an issue that remains largely unexplored. He...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.94.15.8036

    authors: Gonzalez-Couto E,Klages N,Strubin M

    更新日期:1997-07-22 00:00:00

  • Role of an esterase in flavor volatile variation within the tomato clade.

    abstract::Tomato flavor is dependent upon a complex mixture of volatiles including multiple acetate esters. Red-fruited species of the tomato clade accumulate a relatively low content of acetate esters in comparison with the green-fruited species. We show that the difference in volatile ester content between the red- and green-...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1216515109

    authors: Goulet C,Mageroy MH,Lam NB,Floystad A,Tieman DM,Klee HJ

    更新日期:2012-11-13 00:00:00

  • Cardiac hypertrophy and sudden death in mice with a genetically clamped renin transgene.

    abstract::Several mouse models have already proved valuable for investigating hypertrophic responses to cardiac stress. Here, we characterize one caused by a well defined single copy transgene, RenTgMK, that genetically clamps plasma renin and thence angiotensin II at high levels. All of the transgenic males develop concentric ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0307333101

    authors: Caron KM,James LR,Kim HS,Knowles J,Uhlir R,Mao L,Hagaman JR,Cascio W,Rockman H,Smithies O

    更新日期:2004-03-02 00:00:00

  • Targeted disruption of the galanin gene reduces the number of sensory neurons and their regenerative capacity.

    abstract::The neuropeptide galanin is expressed developmentally in the dorsal root ganglion (DRG) and is rapidly up-regulated 120-fold after peripheral nerve section in the adult. Here we report that adult mice carrying a loss-of-function mutation in the galanin gene have a 13% reduction in the number of cells in the DRG associ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.210221897

    authors: Holmes FE,Mahoney S,King VR,Bacon A,Kerr NC,Pachnis V,Curtis R,Priestley JV,Wynick D

    更新日期:2000-10-10 00:00:00

  • Simultaneous synthesis of types I and III collagen by fibroblasts in culture.

    abstract::Specific antibodies against types I and III collagens and procollagens were used to localize these proteins in cultured human cells. These studies indicate that the same cell makes both proteins. No type III procollagen synthesis was observed in cells from two patients with two patients with the Ehlers-Danlos type IV ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.11.4037

    authors: Gay S,Martin GR,Muller PK,Timpl R,Kuhn K

    更新日期:1976-11-01 00:00:00