Protein Phosphorylation in Signal Transduction and Disease

Total Page:16

File Type:pdf, Size:1020Kb

Protein Phosphorylation in Signal Transduction and Disease News A Special Symposium to celebrate Philip Cohen’s 21 years as Director of the MRC Protein Phosphorylation Unit, 24–26 June 2012, University of Dundee Protein Phosphorylation in Signal Transduction and Disease Peter Shepherd (University of Auckland) Downloaded from http://portlandpress.com/biochemist/article-pdf/34/4/59/5271/bio034040059.pdf by guest on 01 October 2021 The Dundee ‘microclimate’ that Sir Philip Cohen has promoted so well over the years with a special award by the Biochemical Journal to was remarkably in evidence as sun greeted more than 200 attendees who had gathered acknowledge the enormous impact of his publications from all over the world to acknowledge his achievements over the last 21 years in in the journal which in themselves have accumulated building the MRC Protein Phosphorylation Unit. more than 10 000 citations and as a body of work have The invitation-only meeting was organized by John Rouse and Dario Alessi, who an h-index of 37. is the new director of the unit, and the invitees featured a range of alumni of Philip’s The scientific proceedings concluded fittingly with a laboratory and the MRC unit as well as people he had collaborated with over the years. presentation from Philip himself who revealed the new The meeting opened with talks from Tim Hunt and Ed Fisher, Philip’s postdoctoral direction for his research now that he has stepped down supervisor in the 1960s. It was no surprise that the subsequent list of speakers comprised as head of the unit. In this presentation he described yet a veritable who’s who of cell signalling researchers whose talks provided an excellent to be published work which provides strong evidence balance of a new results interwoven with historical perspective. There were also talks that the SIK family of kinases play an important in from some of Philip’s collaborators in industry recounting the history of collaborations regulating innate immunity. This looks set to be yet in molecular screening and antibody production that have provided extremely valuable another major contribution to the field of cell signalling resources for not only academia, but also for the drug discovery industry. and who would doubt that Philip will continue to make Naturally, the meeting focused on the impact Philip’s work has made, and many such contributions in the future. In keeping with his h-index of 124 speaks for itself. Many speakers also commented on Philip’s Philip’s style, the meeting concluded with a dinner in contributions to the University of Dundee and Dundee itself and also to his the spectacular dining room of Glamis Castle, a fitting long-term support for European scientific institutions including the Biochemical end to a meeting to honour someone who never does Society, of which he is a past president. During the meeting, Philip was presented things in half measure. ■ Sir Philip Cohen with Professor Chris Marshall from the Institute of Cancer Research Sir Philip Cohen with Eddy Fischer (his postdoctoral London at Glamis Castle supervisor from Seattle) at Glamis Castle August 2012 © Biochemical Society 59.
Recommended publications
  • Tyrosine Kinase – Role and Significance in Cancer
    Int. J. Med. Sci. 2004 1(2): 101-115 101 International Journal of Medical Sciences ISSN 1449-1907 www.medsci.org 2004 1(2):101-115 ©2004 Ivyspring International Publisher. All rights reserved Review Tyrosine kinase – Role and significance in Cancer Received: 2004.3.30 Accepted: 2004.5.15 Manash K. Paul and Anup K. Mukhopadhyay Published:2004.6.01 Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Sector-67, S.A.S Nagar, Mohali, Punjab, India-160062 Abstract Tyrosine kinases are important mediators of the signaling cascade, determining key roles in diverse biological processes like growth, differentiation, metabolism and apoptosis in response to external and internal stimuli. Recent advances have implicated the role of tyrosine kinases in the pathophysiology of cancer. Though their activity is tightly regulated in normal cells, they may acquire transforming functions due to mutation(s), overexpression and autocrine paracrine stimulation, leading to malignancy. Constitutive oncogenic activation in cancer cells can be blocked by selective tyrosine kinase inhibitors and thus considered as a promising approach for innovative genome based therapeutics. The modes of oncogenic activation and the different approaches for tyrosine kinase inhibition, like small molecule inhibitors, monoclonal antibodies, heat shock proteins, immunoconjugates, antisense and peptide drugs are reviewed in light of the important molecules. As angiogenesis is a major event in cancer growth and proliferation, tyrosine kinase inhibitors as a target for anti-angiogenesis can be aptly applied as a new mode of cancer therapy. The review concludes with a discussion on the application of modern techniques and knowledge of the kinome as means to gear up the tyrosine kinase drug discovery process.
    [Show full text]
  • Principles of Protein Phosphorylation Biophysical Chemistry 1, Fall 2010
    Principles of protein phosphorylation Biophysical Chemistry 1, Fall 2010 Signalling “cascades” Structural biology of phosphorylation Web assignment: http://pkr.genomics.purdue.edu Reversible protein phosphorylation Enzymatic reaction Posttranslational Control ΔG~12kcal/mol Kinases and phosphatases Phosphatase Kinase dephospho- phosphorylates rylates General Examples Signalling overview Cells are way too complex! MAPK/ERK Signaling Pathway RousExample:sarcoma virus Rous (RSV) sarcoma virus (RSV) gag - encodes capsid proteins pol - encodes reverse transcriptase env - encodes envelope proteins src - encodes a tyrosine kinase that attaches phosphate groups to the amino acid tyrosine in host cell proteins MutationsExample:, viruses Rous and cancer sarcoma virus (RSV) v-src lacks the C-terminal inhibitory phosphorylation site (tyrosine-527), and is therefore constitutively active as opposed to normal src (c-src) Continuous cell profileration tumor BiophysicsStructural of signalling Effect of Phosphorylation Phosphorylation is an important regulatory mechanism Can reversible attach/detach a phosphate and therefore switch “on”/”off” the function Effect of phosphorylation is manifold • Conformational change • Ordering/disordering • Electrostatic effects • Alternate binding behavior Signalling by reorientation Reorientation: A conformational switch Rmsd = 2.5Å DHP (red) and Z-Score = 4.6 DHPs74e (blue) (>3.6 same fold) Title: A phosphorylation-induced conformation change in dematin headpiece Author(s): Jiang ZHG, McKnight CJ Source: STRUCTURE Volume: 14 Issue: 2 Pages: 379-387 Published: FEB 2006 Order/disorderDisordering:transitions NtrC, a molecular switch upon phosphorylation Orange-yellow: unphosphorylated NtrC blue-cyan: phosphorylated NtrC Volkman et al., Science 2001, 291, 2429-33 Alternate Binding: SRC SH2 Src/SH2 interactions: binding vs release domain binding ExpectedCan conformationalwe understand effects (predict) the effect of phosphorylation Electrostatics Hydrogen bonding Size Mutational analysis Experimental data Lubman, O.Y.
    [Show full text]
  • Protein Kinases Phosphorylation/Dephosphorylation Protein Phosphorylation Is One of the Most Important Mechanisms of Cellular Re
    Protein Kinases Phosphorylation/dephosphorylation Protein phosphorylation is one of the most important mechanisms of cellular responses to growth, stress metabolic and hormonal environmental changes. Most mammalian protein kinases have highly a homologous 30 to 32 kDa catalytic domain. • Most common method of reversible modification - activation and localization • Up to 1/3 of cellular proteins can be phosphorylated • Leads to a very fast response to cellular stress, hormonal changes, learning processes, transcription regulation .... • Different than allosteric or Michealis Menten regulation Protein Kinome To date – 518 human kinases known • 50 kinase families between yeast, invertebrate and mammaliane kinomes • 518 human PKs, most (478) belong to single super family whose catalytic domain are homologous. • Kinase dendrogram displays relative similarities based on catalytic domains. • AGC (PKA, PKG, PKC) • CAMK (Casein kinase 1) • CMGC (CDC, MAPK, GSK3, CLK) • STE (Sterile 7, 11 & 20 kinases) • TK (Tryosine kinases memb and cyto) • TKL (Tyrosine kinase-like) • Phosphorylation stabilized thermodynamically - only half available energy used in adding phosphoryl to protein - change in free energy forces phosphorylation reaction in one direction • Phosphatases reverse direction • The rate of reaction of most phosphatases are 1000 times faster • Phosphorylation occurs on Ser/The or Tyr • What differences occur due to the addition of a phosphoryl group? • Regulation of protein phosphorylation varies depending on protein - some turned on or off
    [Show full text]
  • Β-Catenin Signaling Dynamics Regulate Cell Fate in Differentiating Neural Stem Cells
    β-Catenin signaling dynamics regulate cell fate in differentiating neural stem cells Alyssa B. Rosenblooma, Marcin Tarczynski a, Nora Lama, Ravi S. Kaneb,1, Lukasz J. Bugaja,c,1, and David V. Schaffera,d,e,f,1 aDepartment of Bioengineering, University of California, Berkeley, CA 94720; bSchool of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332; cDepartment of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104; dDepartment of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720; eDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720; and fHelen Wills Neuroscience Institute, University of California, Berkeley, CA 94720 Edited by Randall T. Moon, University of Washington, Seattle, WA, and approved September 21, 2020 (received for review May 4, 2020) Stem cells undergo differentiation in complex and dynamic environ- of how signaling dynamics impact cellular function. Optogenetics ments wherein instructive signals fluctuate on various timescales. has recently emerged as a field in which light—which can readily Thus, cells must be equipped to properly respond to the timing of be varied in intensity, space, and time—is harnessed to precisely signals, for example, to distinguish sustained signaling from transient modulate cell-signaling pathways. In this approach, light-sensitive noise. However, how stem cells respond to dynamic variations in proteins are engineered to interface with specific signaling path- differentiation cues is not well characterized. Here, we use optoge- ways, and the subsequent introduction of such an optogenetic netic activation of β-catenin signaling to probe the dynamic responses construct into cells renders the signaling pathway responsive to of differentiating adult neural stem cells (NSCs).
    [Show full text]
  • Structural Basis of O-Glcnac Recognition by Mammalian 14-3-3 Proteins
    Structural basis of O-GlcNAc recognition by mammalian 14-3-3 proteins Clifford A. Tolemana,1, Maria A. Schumachera,1, Seok-Ho Yub, Wenjie Zenga, Nathan J. Coxa, Timothy J. Smitha, Erik J. Soderblomc, Amberlyn M. Wandsb, Jennifer J. Kohlerb, and Michael Boycea,2 aDepartment of Biochemistry, Duke University School of Medicine, Durham, NC 27710; bDepartment of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390; and cDuke Proteomics and Metabolomics Core Facility, Center for Genomic and Computational Biology, Duke University, Durham, NC 27710 Edited by Carolyn R. Bertozzi, Stanford University, Stanford, CA, and approved April 23, 2018 (received for review December 24, 2017) O-GlcNAc is an intracellular posttranslational modification that gov- Results erns myriad cell biological processes and is dysregulated in human We developed a biochemical approach to test the hypothesis that diseases. Despite this broad pathophysiological significance, the O-GlcNAc is specifically recognized by mammalian reader pro- biochemical effects of most O-GlcNAcylation events remain unchar- teins. First, we derived a consensus O-GlcNAcylated peptide acterized. One prevalent hypothesis is that O-GlcNAc moieties may sequence by aligning 802 mapped Ser-O-GlcNAc sites (34–36) be recognized by “reader” proteins to effect downstream signaling. (Fig. 1A)(www.phosphosite.org). We noted that a Pro-Val-Ser However, no general O-GlcNAc readers have been identified, leav- tripeptide observed previously in smaller datasets (37, 38) also ing a considerable gap in the field. To elucidate O-GlcNAc signaling emerged in our sequence, suggesting that this motif may be mechanisms, we devised a biochemical screen for candidate O-GlcNAc important for O-GlcNAc modification and/or recognition.
    [Show full text]
  • The Origins of Protein Phosphorylation
    historical perspective The origins of protein phosphorylation Philip Cohen The reversible phosphorylation of proteins is central to the regulation of most aspects of cell func- tion but, even after the first protein kinase was identified, the general significance of this discovery was slow to be appreciated. Here I review the discovery of protein phosphorylation and give a per- sonal view of the key findings that have helped to shape the field as we know it today. he days when protein phosphorylation was an abstruse backwater, best talked Tabout between consenting adults in private, are over. My colleagues no longer cringe on hearing that “phosphorylase kinase phosphorylates phosphorylase” and their eyes no longer glaze over when a “”kinase kinase kinase” is mentioned. This is because protein phosphorylation has gradu- ally become an integral part of all the sys- tems they are studying themselves. Indeed it would be difficult to find anyone today who would disagree with the statement that “the reversible phosphorylation of proteins regu- lates nearly every aspect of cell life”. Phosphorylation and dephosphorylation, catalysed by protein kinases and protein phosphatases, can modify the function of a protein in almost every conceivable way; for Carl and Gerty Cori, the 1947 Nobel Laureates. Picture: Science Photo Library. example by increasing or decreasing its bio- logical activity, by stabilizing it or marking it for destruction, by facilitating or inhibiting movement between subcellular compart- so long before its general significance liver enzyme that catalysed the phosphory- ments, or by initiating or disrupting pro- was appreciated? lation of casein3. Soon after, Fischer and tein–protein interactions.
    [Show full text]
  • Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia
    cancers Review Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia Kalpana K. Bhanumathy 1,*, Amrutha Balagopal 1, Frederick S. Vizeacoumar 2 , Franco J. Vizeacoumar 1,3, Andrew Freywald 2 and Vincenzo Giambra 4,* 1 Division of Oncology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (A.B.); [email protected] (F.J.V.) 2 Department of Pathology and Laboratory Medicine, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (F.S.V.); [email protected] (A.F.) 3 Cancer Research Department, Saskatchewan Cancer Agency, 107 Wiggins Road, Saskatoon, SK S7N 5E5, Canada 4 Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS Casa Sollievo della Sofferenza, 71013 San Giovanni Rotondo, FG, Italy * Correspondence: [email protected] (K.K.B.); [email protected] (V.G.); Tel.: +1-(306)-716-7456 (K.K.B.); +39-0882-416574 (V.G.) Simple Summary: Protein phosphorylation is a key regulatory mechanism that controls a wide variety of cellular responses. This process is catalysed by the members of the protein kinase su- perfamily that are classified into two main families based on their ability to phosphorylate either tyrosine or serine and threonine residues in their substrates. Massive research efforts have been invested in dissecting the functions of tyrosine kinases, revealing their importance in the initiation and progression of human malignancies. Based on these investigations, numerous tyrosine kinase inhibitors have been included in clinical protocols and proved to be effective in targeted therapies for various haematological malignancies.
    [Show full text]
  • The T-Cell Protein Tyrosine Phosphatase Is Phosphorylated On
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Cold Spring Harbor Laboratory Institutional Repository Biochem. J. (2004) 380, 939–949 (Printed in Great Britain) 939 The T-cell protein tyrosine phosphatase is phosphorylated on Ser-304 by cyclin-dependent protein kinases in mitosis Patricia BUKCZYNSKA*1, Manuela KLINGLER-HOFFMANN*1, Kenneth I. MITCHELHILL†2, Mark H. C. LAM*3, Melissa CICCOMANCINI*4, Nicholas K. TONKS‡, Boris SARCEVIC§, Bruce E. KEMP† and Tony TIGANIS*5 *Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Victoria 3800, Australia, †St. Vincent’s Institute of Medical Research, Melbourne, Victoria 3065, Australia, ‡Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, U.S.A., and §Cancer Research Program, Garvan Institute of Medical Research, Darlinghurst, New South Wales 2010, Australia Two alternatively spliced forms of the human protein tyrosine no apparent effect on TC45 localization. Ser-304 phosphorylation phosphatase TCPTP (T-cell protein tyrosine phosphatase) exist: was ablated when cells were treated with the CDK (cyclin- a 48 kDa form that is targeted to the endoplasmic reticulum dependent protein kinase) inhibitors roscovitine or SU9516, (TC48) and a shorter 45 kDa form that is targeted to the but remained unaltered when ERK1/2 activation was inhibited nucleus (TC45). In this study we have identified Ser-304 (Phe301- with the MEK (mitogen-activated protein kinase/extracellular- Asp-His-Ser304-Pro-Asn-Lys307) as a major TCPTP phosphory- signal-regulated kinase kinase) inhibitor PD98059. In addition, lation site and demonstrate that TC45, but not TC48, is phos- recombinant CDKs, but not the Polo-like kinase Plk1, phos- phorylated on this site in vivo.
    [Show full text]
  • Stability of Phosphoprotein As a Biological Marker of Tumor Signaling Amanda F
    Imaging, Diagnosis, Prognosis Stability of Phosphoprotein as a Biological Marker of Tumor Signaling Amanda F. Baker,1Tomislav Dragovich,1Nathan T.Ihle,1RyanWilliams,1 Cecilia Fenoglio-Preiser,2 and Garth Powis1 Abstract Purpose:The purpose of the study was to evaluate the stability of phosphoprotein as a marker of signaling activity in human tumors using clinical samples and xenografts. Experimental Design: The expression of phospho-Ser473-Akt (p-Akt) was assessed by immunohistochemistry in paraffin-embedded samples from patients enrolled in a Southwest Oncology Group clinical trial of gastroesophageal junction tumors and by immunohistochemistry and Western blotting in human colon tumor xenografts at various times after removal from the animal. Results: Clinical samples had evaluable p-Akt staining only when obtained as biopsies (9 of 13) and no staining was observed in tumors obtained as surgically resected samples (0 of 15). In HT- 29 colon cancer xenografts, p-Akt staining was present in fresh sample but not in tissue that had been allowed to stand for 30 minutes at room temperature.Western blotting of HT-29 tumor xen- ografts at room temperature showed a slow decrease in total Akt with a half-life of 180 minutes and a rapid decrease in p-Akt with a half-life of 20 minutes. Conclusions: Caution should be used when using phosphoprotein levels in human tumor speci- mens to measure intrinsic signaling activity or drug effects because of the potential for rapid de- phosphorylation. Rapid processing of biopsies is essential and postoperative surgical samples may be of limited value because of the time to fixation.
    [Show full text]
  • Reversible Phosphorylation of Rpn1 Regulates 26S Proteasome Assembly and Function
    Reversible phosphorylation of Rpn1 regulates 26S proteasome assembly and function Xiaoyan Liua,1, Weidi Xiaob,1, Yanan Zhanga, Sandra E. Wileyc, Tao Zuob, Yingying Zhenga, Natalie Chenc, Lu Chena, Xiaorong Wangd,e, Yawen Zhenga, Lan Huangd,e, Shixian Lina, Anne N. Murphyc, Jack E. Dixonc,f,g, Ping Xub, and Xing Guoa,2 aLife Sciences Institute, Zhejiang University, Hangzhou, P. R. China 310058; bState Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Institute of Lifeomics, Beijing, P. R. China 102206; cDepartment of Pharmacology, University of California San Diego, La Jolla, CA 92093; dDepartment of Physiology and Biophysics, University of California, Irvine, CA 92697; eDepartment of Developmental and Cell Biology, University of California, Irvine, CA 92697; fDepartment of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093; and gDepartment of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093 Edited by Alfred L. Goldberg, Harvard Medical School, Boston, MA, and approved November 21, 2019 (received for review July 22, 2019) The fundamental importance of the 26S proteasome in health and each containing a pair of Rpt subunits, namely Rpt1-Rpt2-Rpn1- disease suggests that its function must be finely controlled, and S5b, Rpt3-Rpt6-p28-PAAF1, and Rpt4-Rpt5-p27, with S5b, p28, yet our knowledge about proteasome regulation remains limited. PAAF1, and p27 being specific chaperone proteins (13–16). Joining Posttranslational modifications, especially phosphorylation, of of these precursor complexes as well as sequential leaving of the proteasome subunits have been shown to impact proteasome chaperones leads to formation of the base, and subsequent in- function through different mechanisms, although the vast major- corporation of the remaining Rpn subunits completes 19S RP ity of proteasome phosphorylation events have not been studied.
    [Show full text]
  • The Crosstalk Between O-Glcnacylation and Phosphorylation in Insulin Signaling
    THE CROSSTALK BETWEEN O-GLCNACYLATION AND PHOSPHORYLATION IN INSULIN SIGNALING by Ping Hu A dissertation submitted to Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland December, 2015 © Ping Hu All Rights Reserved ABSTRACT Extensive crosstalk between phosphorylation and O-GlcNAcylation is involved in regulating insulin signaling. In addition, many of the key regulators of insulin signaling have been shown to be O-GlcNAcylated. In this study, we first explored the crosstalk between phosphorylation and O-GlcNAcylation globally, and then focused on an upstream signaling regulator key to insulin signaling, insulin receptor substrate 1, IRS1. Through regulating cellular O-GlcNAcylation levels in differentiated adipocyte 3T3-L1 cells by specific OGT and OGA inhibitors, we measured the affects of O-GlcNAcylation on the phosphorylation status of key molecules. We observed that O-GlcNAcylation regulates the flux through the two main divergent cellular pathways of insulin signaling, one regulating glucose metabolism and the other regulating cellular proliferation via MAP kinase. Decreased O-GlcNAcylation was associated with increased glucose utilization, and increased O-GlcNAcylation was associated with increased insulin-induced cellular proliferation pathways. Since IRS1 is early in the insulin signaling pathway and is both highly O-GlcNAcylated and highly phosphorylated, we focused on the crosstalk of those two PTMs on IRS1. The phosphorylation spectrum of IRS1 revealed by 2D electrophoretic analyses, partially explained why decreased O-GlcNAcylation facilitated glucose uptake. Two new O- GlcNAcylation sites were mapped through CID/ETD mass spectrometry. With help of protein truncation and site mutagenesis techniques, we propose a mechanism suggesting that the O-GlcNAcylation of Ser635, may be a negative regulator of the C-terminal O- GlcNAcylations on IRS1.
    [Show full text]
  • Phosphoproteomics for the Masses
    REVIEW Phosphoproteomics for the Masses Paul A. Grimsrud†, Danielle L. Swaney†, Craig D. Wenger†, Nicole A. Beauchene‡, and Joshua J. Coon†,‡,* †Departments of Chemistry and ‡Biomolecular Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706 PHOSPHOPROTEOMICS APPLICATIONS Protein phosphorylation is a central mechanism of sig- ABSTRACT Protein phosphorylation serves as a primary mechanism of signal nal transduction across species, with kinases and phos- transduction in the cells of biological organisms. Technical advancements over the phatases accounting for 2Ϫ4% of eukaryotic proteomes last several years in mass spectrometry now allow for the large-scale identifica- (1, 2). Current estimates suggest that one-third of eu- tion and quantitation of in vivo phosphorylation at unprecedented levels. These de- karyotic proteins are phosphorylated (3, 4); determin- velopments have occurred in the areas of sample preparation, instrumentation, ing the sites, abundances, and roles of each of these quantitative methodology, and informatics so that today, 10 000Ϫ20 000 phos- modifications in a biological sample is a critical chal- phorylation sites can be identified and quantified within a few weeks. With the lenge. Traditional biochemical techniques are chiefly rapid development and widespread availability of such data, its translation into limited to testing how phosphoryl modifications at spe- biological insight and knowledge is a current obstacle. Here we present an over- cific sites affect a single protein of interest (5). Identifica- view of how this technology came to be and is currently applied, as well as future tion of unknown in vivo phosphoryl modification sites challenges for the field. on a broad scale, however, is simply not possible with these approaches.
    [Show full text]