2019 Annual Report
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Realizing the Allosteric Potential of the Tetrameric Protein Kinase a RIΑ Holoenzyme
Structure Article Realizing the Allosteric Potential of the Tetrameric Protein Kinase A RIa Holoenzyme Angela J. Boettcher,1,6 Jian Wu,1,6 Choel Kim,2 Jie Yang,1 Jessica Bruystens,1 Nikki Cheung,1 Juniper K. Pennypacker,1,3 Donald A. Blumenthal,4 Alexandr P. Kornev,3,5 and Susan S. Taylor1,3,5,* 1Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, CA 92093, USA 2Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA 3Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093, USA 4Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112, USA 5Howard Hughes Medical Institute, University of California at San Diego, La Jolla, CA 92093, USA 6These authors contributed equally to this work *Correspondence: [email protected] DOI 10.1016/j.str.2010.12.005 SUMMARY the active site cleft in the C subunit in the inactive holoenzyme but is disordered in the dissociated free R subunits (Li et al., PKA holoenzymes containing two catalytic (C) 2000). The linker, as summarized in Figure 1, can be divided subunits and a regulatory (R) subunit dimer are acti- into three segments, the consensus inhibitor site (P-3 to P+1), vated cooperatively by cAMP. While cooperativity the N-linker that joins the inhibitor site to the D/D domain, and involves the two tandem cAMP binding domains in the C-linker that becomes ordered in the heterodimeric holoen- each R-subunit, additional cooperativity is associ- zyme complex. While much has been learned from the structures ated with the tetramer. -
A Perspective on Mechanisms of Protein Tetramer Formation
Biophysical Journal Volume 85 December 2003 3587–3599 3587 A Perspective on Mechanisms of Protein Tetramer Formation Evan T. Powers* and David L. Powersy *Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037; and yDepartment of Mathematics and Computer Science, Clarkson University, Potsdam, New York 13699 ABSTRACT Homotetrameric proteins can assemble by several different pathways, but have only been observed to use one, in which two monomers associate to form a homodimer, and then two homodimers associate to form a homotetramer. To determine why this pathway should be so uniformly dominant, we have modeled the kinetics of tetramerization for the possible pathways as a function of the rate constants for each step. We have found that competition with the other pathways, in which homotetramers can be formed either by the association of two different types of homodimers or by the successive addition of monomers to homodimers and homotrimers, can cause substantial amounts of protein to be trapped as intermediates of the assembly pathway. We propose that this could lead to undesirable consequences for an organism, and that selective pressure may have caused homotetrameric proteins to evolve to assemble by a single pathway. INTRODUCTION Many proteins must be homotetrameric to be functional. ‘‘MDT’’ stands for monomer-dimer-tetramer and ‘‘a’’ and Prominent examples include transcription factors (e.g., p53) ‘‘b’’ indicate the type of homodimer formed. The pathways (Friedman et al., 1993), transport proteins (e.g., trans- that include monomers, homodimers, homotrimers, and thyretin) (Blake et al., 1974), potassium channels (Deutsch, homotetramers will be denoted MDRT, which stands for 2002; Miller, 2000), water channels (Fujiyoshi et al., 2002), monomer-dimer-trimer-tetramer. -
HOOK™ Maleimide Activated Streptavidin for Conjugation of Streptavidin to Sulfhydryl Groups Containing Proteins, Peptides and Ligands
G-Biosciences 1-800-628-7730 1-314-991-6034 [email protected] A Geno Technology, Inc. (USA) brand name HOOK™ Maleimide Activated Streptavidin For conjugation of Streptavidin to sulfhydryl groups containing proteins, peptides and ligands (Cat. #786-1653, 786-1654) think proteins! think G-Biosciences www.GBiosciences.com INTRODUCTION ................................................................................................................. 3 ITEMS SUPPLIED ................................................................................................................ 4 STORAGE CONDITIONS ...................................................................................................... 4 ADDITIONAL ITEMS NEEDED .............................................................................................. 4 IMPORTANT INFORMATION .............................................................................................. 4 PROTOCOL ......................................................................................................................... 4 PREPARATION OF PROTEIN FOR CONJUGATION TO MALEIMIDE ACTIVATED PROTEIN 4 CONJUGATION REACTION ............................................................................................. 5 STORAGE OF CONJUGATED ANTIBODIES/PROTEINS ......................................................... 5 RELATED PRODUCTS .......................................................................................................... 5 Page 2 of 6 INTRODUCTION Streptavidin is a non-glycosylated -
Vaccine Hesitancy
WHY CHILDREN WORKSHOP ON IMMUNIZATIONS ARE NOT VACCINATED? VACCINE HESITANCY José Esparza MD, PhD - Adjunct Professor, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, USA - Robert Koch Fellow, Robert Koch Institute, Berlin, Germany - Senior Advisor, Global Virus Network, Baltimore, MD, USA. Formerly: - Bill & Melinda Gates Foundation, Seattle, WA, USA - World Health Organization, Geneva, Switzerland The value of vaccination “The impact of vaccination on the health of the world’s people is hard to exaggerate. With the exception of safe water, no other modality has had such a major effect on mortality reduction and population growth” Stanley Plotkin (2013) VACCINES VAILABLE TO PROTECT AGAINST MORE DISEASES (US) BASIC VACCINES RECOMMENDED BY WHO For all: BCG, hepatitis B, polio, DTP, Hib, Pneumococcal (conjugated), rotavirus, measles, rubella, HPV. For certain regions: Japanese encephalitis, yellow fever, tick-borne encephalitis. For some high-risk populations: typhoid, cholera, meningococcal, hepatitis A, rabies. For certain immunization programs: mumps, influenza Vaccines save millions of lives annually, worldwide WHAT THE WORLD HAS ACHIEVED: 40 YEARS OF INCREASING REACH OF BASIC VACCINES “Bill Gates Chart” 17 M GAVI 5.6 M 4.2 M Today (ca 2015): <5% of children in GAVI countries fully immunised with the 11 WHO- recommended vaccines Seth Berkley (GAVI) The goal: 50% of children in GAVI countries fully immunised by 2020 Seth Berkley (GAVI) The current world immunization efforts are achieving: • Equity between high and low-income countries • Bringing the power of vaccines to even the world’s poorest countries • Reducing morbidity and mortality in developing countries • Eliminating and eradicating disease WHY CHILDREN ARE NOT VACCINATED? •Vaccines are not available •Deficient health care systems •Poverty •Vaccine hesitancy (reticencia a la vacunacion) VACCINE HESITANCE: WHO DEFINITION “Vaccine hesitancy refers to delay in acceptance or refusal of vaccines despite availability of vaccination services. -
Clinical Scholars Visit Amish Research Clinic in Lancaster County, PA by Patrick Brunner, MD Dr
Spring 2018 Center for Clinical and Translational Science e- e-NewsletterNewsletter Center News James Krueger and Marina Caskey, Representing the Nussenzweig Lab Team, Honored at Translational Science 2018 Meeting By Hospital Leadership which fulfill the translation research Dr. James Krueger received paradigm of going from bench to the Association for Clinical and bedside, have been recognized by this Translational Science (ACTS) most prestigious national recognition. Distinguished Investigator Award for his It is well deserved!” Upon receiving the groundbreaking research on psoriasis at award, Dr. Krueger noted, “This award the Translational Science 2018 meeting would not have been possible without attended by more than 1,100 people in the support of many others associated Washington, DC in April. His research with the Rockefeller CTSA enterprise: has led to a fundamental change in my lab members, the nursing staff, the paradigm for understanding the all other support departments and, pathophysiology of the disorder, and of course, hundreds of patients who this in turn has led to the development directly tested progressively better drugs of a series of novel medications that that are now used to so effectively treat precisely modulate the immune system psoriasis.” and dramatically improve the therapy of the disorder. Each year, Clinical Research Forum sponsors a competition to identify Dr. Caskey Receiving “Top Ten” Award from the “Top Ten” Clinical Research Drs. Harry Selker and Herb Pardes studies reported in the previous year. commented that “The study reflects the The competition is intense and so it very best in translational science: the is a true tribute to the novelty and careful analysis of patient phenotypes; importance of the study led by Dr. -
Report on Global Surveillance of Epidemic-Prone Infectious Diseases, 2000
Pan American Health Organization Regional Office of the World Health Organization PAHO/DPC/CD-V/243/03 Original: English Report: Workshop on Dengue Burden Studies (Washington, DC, 5-7 November 2002) Convened by The Pan American Health Organization The Rockefeller Foundation The Pediatric Dengue Vaccine Initiative Executive Summary Background Dengue fever (DF) and dengue hemorrhagic fever (DHF) are caused by the mosquito borne virus, dengue virus, of which there are four antigenically distinct serotypes. It is estimated that annually these viruses cause at least 20 million infections worldwide leading to some 24,000 deaths (WHO, http://www.who.int/health_topics/dengue/en/ ). The alarming rise in dengue hemorrhagic fever in the world today is illustrated most starkly by the chart below which represents data from the World Health Organization (WHO) showing the rise of DHF cases over the last four decades. Indeed the first two years of the new millennium has seen outbreak after outbreak of DHF not only in Southeast Asia where DHF has been seen for half a century, but also in many countries of South and Central America. REPORTED CASES OF DHF 600 ds 500 housan t 400 n d i e 300 200 r report e 100 numb 0 1955-1959 1960-1069 1970-1979 1980-1989 1990-1998 Source: WHO; adapted from http://www.who.int/health_topics/dengue/en/ While there is no doubt that severe dengue is spreading from countries in Southeast Asia to countries in the Pacific and in the Americas, there is also no doubt that many international efforts into the development of dengue vaccines have led to a number of promising vaccine candidates which may offer some solutions to the control of this disease. -
V21 MTG 0033 List of Particip
THE FIRST WHO INTEGRATED MEETING ON DEVELOPMENT AND CLINICAL TRIALS OF INFLUENZA VACCINES THAT INDUCE BROADLY PROTECTIVE AND LONG-LASTING IMMUNE RESPONSES Baptist University, Hong Kong 24 – 26 January 2013 LIST OF PARTICIPANTS Dr Akira Ainai, Emeritus Member, National Institute of Infectious Diseases, Tokyo, Japan Dr Christopher Ambrose, Senior Director, Medical Affairs, MedImmune, Gaithersburg, United States of America Professor Filippo Ansaldi, Health Sciences, University of Genoa and Hygiene Unit, Genoa, Italy Dr Robert L. Atmar, Associate Professor, Department of Medicine and Molecular Virology & Microbiology, Baylor College of Medicine, Houston, United States of America Dr Emma Ball, CSL Limited A.C.N., Parkville, Victoria, Australia Dr Pamuk Bilsel, FluGen Inc., Madison, United States of America Mr Albert Chan, President, Hong Kong Baptist University, Hong Kong, China Dr Paul Chan, Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China Dr Marta Coehlo Nunes, Respiratory and Meningeal Pathogens Research Unit, Wits University, Johannesburg, South Africa Dr Ben Cowling, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China Dr Manon Cox, President and CEO, Protein Sciences Corporation, Meridien, United States of America Professor Rebecca Cox, Influenza Centre, The Gade Institute, University of Bergen, Bergen, 5021 Norway Dr Giuseppe Del Giudice, Global Head Translation Medicine, Novartis Vaccines and Diagnostics S.p.A, Divisione Biologici -
5Th Annual Short Course in Medical Virology Baltimore, MD August 5-11
5th Annual Short Course in Medical Virology Baltimore, MD ǀ August 5-11, 2018 ǀ Program Locations: IHV ǀ Baltimore JH ǀ Baltimore NIH ǀ Bethesda Sunday, August 5 3:00 PM Hotel Check In 6:00-8:00 PM Short Course Opening Dinner at Diamond Tavern Monday, August 6 9:00-10:00 AM Shyam Kottilil, MD, PhD, Institute of Human Virology "Hepatitis C" 10:00-10:15 AM Coffee break 10:15- 11:15 AM Yutaka Tagaya, MD, PhD, Institute of Human Virology "Human T-cell leukemia virus-1, an exceptionally oncogenic retrovirus" 11:15-12:15 PM Robert Gallo, MD, Institute of Human Virology "Human Retroviruses: HTLV and HIV: an Overview” 12:15- 1:15 PM group picture & lunch 1:15-2:15 PM Jose Esparza, MD, PhD, Institute of Human Virology "Vaccines" 2:15-3:15 PM Alfredo Garzino-Demo, PhD, Institute of Human Virology "HIV Pathogenesis" 3:15-3:30 PM Coffee break 3:30- 4:30 PM Neil Constantine, PhD, Insitute of Human Virology "Laboratory diagnostics" Tuesday, August 7 8:15 AM Transportation to Johns Hopkins Bloomberg School of Public Health 9:00-10:00 AM Diane Griffin, PhD, MD, Johns Hopkins Bloomberg School of Public Health "Measles" 10:00-11:00 AM Ken Olson, PhD, Colorado State University "Arboviruses" 11:00-11:30 AM Break 11:30-12:30 AM Marcelo Jacobs-Lorena, PhD, Johns Hopkins Bloomberg School of Public Health Insectary Tour 12:30-1:30 PM Lunch 1:30-2:30 PM Transportation to IHV/ Coffee Break 2:30-4:30 PM GVN Short Course Partcipant's research presentations 4:30-5:30 PM Yuki Furuse, PhD, MD, Tohoku University, Japan New Emerging leader presentation: What can we do and what should we do during Ebola outbreak? Wednesday, August 8 9:00-10:00 AM Ab Osterhaus, DVM, PhD, University of Veterninary Medicine Hannover "One Health" 10:00-11:00 AM Stefan Sarafianos, PhD, Emory School of Medicine "Antiviral drug discovery" 11:30- 12:30 AM Robert Garry, PhD, Tulane University "Ebola and Lassa fever" 12:30-1:30 PM Lunch 1:30-2:30PM Konstantin Chumakov, FDA “Polio and other Enteroviruses” 2:30-3:30 PM Richard H. -
José Esparza MD, Phd - Adjunct Professor, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, USA
Vaccination: lost opportunities José Esparza MD, PhD - Adjunct Professor, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, USA. - Formerly at the World Health Organization (Geneva, Switzerland) and the Bill & Melinda Gates Foundation (Seattle, WA, USA). Outline • The public health value of vaccination • Different types of vaccine hesitancy • Some historical examples of vaccine introduction and impact • The case of dengue vaccines • Conclusions Ten Great Public Health Achievements of the 20th Century (in the US) 1. Vaccines 2. Motor vehicle safety 3. Workplace safety 4. Control of infectious diseases 5. Decline in deaths from hearth disease and stroke 6. Safer and healthier foods 7. Healthier mothers and babies 8. Family planning 9. Fluoridation of drinking water 10. Tobacco as a health hazard www.cdc.gov/about/history/tengpha.htm The value of vaccination “The impact of vaccination on the health of the world’s people is hard to exaggerate. With the exception of safe water, no other modality has had such a major effect on mortality reduction and population growth” Stanley Plotkin (2013) Vaccine-preventable diseases, by year of vaccine development or licensure. United States,1798-1998. Vaccine Introduction Vaccine Introduction Smallpox 1798 Rubella 1969 Rabies 1885 Antrax 1970 Typhoid 1896 Meningitis 1975 Cholera 1896 Pneumonia 1977 Plague 1897 Adenovirus 1980 Diphtheria 1923 Hepatitis B 1981 Pertussis 1926 H. Influenza B 1985 Tetanus 1927 Japanese encephalitis 1992 Tuberculosis 1927 Hepatitis A 1995 -
Modulation of Function, Structure and Clustering of K Channels by Lipids: Lessons Learnt from Kcsa
International Journal of Molecular Sciences Review Modulation of Function, Structure and Clustering of K+ Channels by Lipids: Lessons Learnt from KcsA María Lourdes Renart 1 , Ana Marcela Giudici 1, Clara Díaz-García 2 , María Luisa Molina 1 , Andrés Morales 3, José M. González-Ros 1,* and José Antonio Poveda 1,* 1 Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE), and Instituto de Biología Molecular y Celular (IBMC), Universidad Miguel Hernández, Elche, E-03202 Alicante, Spain; [email protected] (M.L.R.); [email protected] (A.M.G.); [email protected] (M.L.M.) 2 iBB-Institute for Bioengineering and Bioscience, Instituto Superior Técnico, Universidade de Lisboa, 1049-001 Lisboa, Portugal; [email protected] 3 Departamento de Fisiología, Genética y Microbiología, Universidad de Alicante, E-03080 Alicante, Spain; [email protected] * Correspondence: [email protected] (J.M.G.-R.); [email protected] (J.A.P.) Received: 5 March 2020; Accepted: 5 April 2020; Published: 7 April 2020 Abstract: KcsA, a prokaryote tetrameric potassium channel, was the first ion channel ever to be structurally solved at high resolution. This, along with the ease of its expression and purification, made KcsA an experimental system of choice to study structure–function relationships in ion channels. In fact, much of our current understanding on how the different channel families operate arises from earlier KcsA information. Being an integral membrane protein, KcsA is also an excellent model to study how lipid–protein and protein–protein interactions within membranes, modulate its activity and structure. In regard to the later, a variety of equilibrium and non-equilibrium methods have been used in a truly multidisciplinary effort to study the effects of lipids on the KcsA channel. -
Piecing Together the HIV Prevention Puzzle AVAC Report 2009 Acknowledgments
Piecing Together the HIV Prevention Puzzle AVAC Report 2009 Acknowledgments AVAC gratefully acknowledges many friends and colleagues in government, industry, academia and the advocacy community from all over the world for their expertise, guidance and advice as we researched and prepared this Report. AVAC Report 2009 was written and edited by AVAC staff, consultants and board members, and coordinated by Emily Bass. We also want to especially thank Sarah Alexander, Dan Barouch, Linda-Gail Bekker, Seth Berkley, Alan Bernstein, Susan Buchbinder, Gabriela Calazans, Ward Cates, Mark Connors, Larry Corey, Paul de Bakker, Guy de Bruyn, Kevin De Cock, Carl Dieffenbach, Kim Dickson, Jose Esparza, Tim Farley, Patricia Fast, Mark Feinberg, Jorge Flores, David Goldstein, Gregg Gonsalves, Glenda Gray, Yasmin Halima, Cate Hankins, Mark Harrington, Barton Haynes, Sharon Hillier, Peggy Johnston, Richard Klausner, Wayne Koff, Katharine Kripke, Jim Kublin, Dave Levin, Udom Likhitwonnawut, Margaret Liu, Ying Ru Lo, Donna Lomangino, Siobhan Malone, Kay Marshall, Betsy Martin, John Mascola, Bonnie Mathieson, Margaret McCluskey, Elizabeth McGrory, James McIntyre, Natasha Mileshina, Lynn Morris, Kevin O’Reilly, Saladin Osmanov, Myra Ozaeta, Giuseppe Pantaleo, Lynn Paxton, Louis Picker, Frances Priddy, Helen Rees, Robert Reinhard, Supachai Rerks-Ngarm, Mike Robertson, Candace Rosen, Nina Russell, Jerry Sadoff, Jeff Safrit, Allan Schultz, Robin Shattock, Guido Silvestri, Joan Tallada, Jim Tartaglia, Gerald Voss, Sabrina Welsh and Carolyn Williamson. AVAC is dedicated -
Multimerization of Homo Sapiens TRPA1 Ion Channel Cytoplasmic Domains
bioRxiv preprint doi: https://doi.org/10.1101/466060; this version posted November 8, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Multimerization of Homo sapiens TRPA1 ion channel cytoplasmic domains 2 Gilbert Q. Martinez, Sharona E. Gordon* 3 Department of Physiology and Biophysics, University of Washington, Seattle, Washington, 4 United States of America 5 6 7 8 9 10 11 12 13 14 15 16 17 *Corresponding author 18 E-mail: [email protected] 19 1 bioRxiv preprint doi: https://doi.org/10.1101/466060; this version posted November 8, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 20 Abstract 21 The transient receptor potential Ankyrin-1 (TRPA1) ion channel is modulated by myriad 22 noxious stimuli that interact with multiple regions of the channel, including cysteine- 23 reactive natural extracts from onion and garlic which modify residues in the cytoplasmic 24 domains. The way in which TRPA1 cytoplasmic domain modification is coupled to opening 25 of the ion-conducting pore has yet to be elucidated. The cryo-EM structure of TRPA1 26 revealed a tetrameric C-terminal coiled-coil surrounded by N-terminal ankyrin repeat 27 domains (ARDs), an architecture shared with the canonical transient receptor potential 28 (TRPC) ion channel family.