The Escherichia Coli Groel Interaction Proteome: Identification and Classification
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Control of Protein Function
3 Control of Protein Function In the cell, precise regulation of protein function is essential to avoid chaos. This chapter describes the most important molecular mechanisms by which protein function is regulated in cells. These range from control of a protein’s location and lifetime within the cell to the binding of regulatory molecules and covalent modifications such as phosphorylation that rapidly switch protein activity on or off. Also covered here are the nucleotide-driven switches in conformation that underlie the action of motor proteins and that regulate many signal transduction pathways. 3-0 Overview: Mechanisms of Regulation 3-1 Protein Interaction Domains 3-2 Regulation by Location 3-3 Control by pH and Redox Environment 3-4 Effector Ligands: Competitive Binding and Cooperativity 3-5 Effector Ligands: Conformational Change and Allostery 3-6 Protein Switches Based on Nucleotide Hydrolysis 3-7 GTPase Switches: Small Signaling G Proteins 3-8 GTPase Switches: Signal Relay by Heterotrimeric GTPases 3-9 GTPase Switches: Protein Synthesis 3-10 Motor Protein Switches 3-11 Regulation by Degradation 3-12 Control of Protein Function by Phosphorylation 3-13 Regulation of Signaling Protein Kinases: Activation Mechanism 3-14 Regulation of Signaling Protein Kinases: Cdk Activation 3-15 Two-Component Signaling Systems in Bacteria 3-16 Control by Proteolysis: Activation of Precursors 3-17 Protein Splicing: Autoproteolysis by Inteins 3-18 Glycosylation 3-19 Protein Targeting by Lipid Modifications 3-20 Methylation, N-acetylation, Sumoylation and Nitrosylation 3-0 Overview: Mechanisms of Regulation Protein function in living cells is precisely regulated A typical bacterial cell contains a total of about 250,000 protein molecules (comprising different amounts of each of several thousand different gene products), which are packed into a volume so small that it has been estimated that, on average, they are separated from one another by a distance that would contain only a few molecules of water. -
Prokaryotic Ubiquitin-Like Protein Remains Intrinsically Disordered When Covalently Attached to Proteasomal Target Proteins Jonas Barandun1,2, Fred F
Barandun et al. BMC Structural Biology (2017) 17:1 DOI 10.1186/s12900-017-0072-1 RESEARCH ARTICLE Open Access Prokaryotic ubiquitin-like protein remains intrinsically disordered when covalently attached to proteasomal target proteins Jonas Barandun1,2, Fred F. Damberger1, Cyrille L. Delley1, Juerg Laederach1, Frédéric H. T. Allain1 and Eilika Weber-Ban1* Abstract Background: The post-translational modification pathway referred to as pupylation marks proteins for proteasomal degradation in Mycobacterium tuberculosis and other actinobacteria by covalently attaching the small protein Pup (prokaryotic ubiquitin-like protein) to target lysine residues. In contrast to the functionally analogous eukaryotic ubiquitin, Pup is intrinsically disordered in its free form. Its unfolded state allows Pup to adopt different structures upon interaction with different binding partners like the Pup ligase PafA and the proteasomal ATPase Mpa. While the disordered behavior of free Pup has been well characterized, it remained unknown whether Pup adopts a distinct structure when attached to a substrate. Results: Using a combination of NMR experiments and biochemical analysis we demonstrate that Pup remains unstructured when ligated to two well-established pupylation substrates targeted for proteasomal degradation in Mycobacterium tuberculosis, malonyl transacylase (FabD) and ketopantoyl hydroxylmethyltransferase (PanB). Isotopically labeled Pup was linked to FabD and PanB by in vitro pupylation to generate homogeneously pupylated substrates suitable for NMR analysis. The single target lysine of PanB was identified by a combination of mass spectroscopy and mutational analysis. Chemical shift comparison between Pup in its free form and ligated to substrate reveals intrinsic disorder of Pup in the conjugate. Conclusion: When linked to the proteasomal substrates FabD and PanB, Pup is unstructured and retains the ability to interact with its different binding partners. -
Yichen – Structure and Allostery of the Chaperonin Groel. Allosteric
Literature Lunch 5-1-13 Yichen – J Mol Biol. 2013 May 13;425(9):1476-87. doi: 10.1016/j.jmb.2012.11.028. Epub 2012 Nov 24. Structure and Allostery of the Chaperonin GroEL. Saibil HR, Fenton WA, Clare DK, Horwich AL. Source Crystallography and Institute of Structural and Molecular Biology, Birkbeck College London, Malet Street, London WC1E 7HX, UK. Abstract Chaperonins are intricate allosteric machines formed of two back-to-back, stacked rings of subunits presenting end cavities lined with hydrophobic binding sites for nonnative polypeptides. Once bound, substrates are subjected to forceful, concerted movements that result in their ejection from the binding surface and simultaneous encapsulation inside a hydrophilic chamber that favors their folding. Here, we review the allosteric machine movements that are choreographed by ATP binding, which triggers concerted tilting and twisting of subunit domains. These movements distort the ring of hydrophobic binding sites and split it apart, potentially unfolding the multiply bound substrate. Then, GroES binding is accompanied by a 100° twist of the binding domains that removes the hydrophobic sites from the cavity lining and forms the folding chamber. ATP hydrolysis is not needed for a single round of binding and encapsulation but is necessary to allow the next round of ATP binding in the opposite ring. It is this remote ATP binding that triggers dismantling of the folding chamber and release of the encapsulated substrate, whether folded or not. The basis for these ordered actions is an elegant system of nested cooperativity of the ATPase machinery. ATP binds to a ring with positive cooperativity, and movements of the interlinked subunit domains are concerted. -
1519038862M28translationand
Paper No. : 15 Molecular Cell Biology Module : 28 Translation and Post-translation Modifications in Eukaryotes Development Team Principal Investigator : Prof. Neeta Sehgal Department of Zoology, University of Delhi Co-Principal Investigator : Prof. D.K. Singh Department of Zoology, University of Delhi Paper Coordinator : Prof. Kuldeep K. Sharma Department of Zoology, University of Jammu Content Writer : Dr. Renu Solanki, Deen Dayal Upadhyaya College Dr. Sudhida Gautam, Hansraj College, University of Delhi Mr. Kiran K. Salam, Hindu College, University of Delhi Content Reviewer : Prof. Rup Lal Department of Zoology, University of Delhi 1 Molecular Genetics ZOOLOGY Translation and Post-translation Modifications in Eukaryotes Description of Module Subject Name ZOOLOGY Paper Name Molecular Cell Biology; Zool 015 Module Name/Title Cell regulatory mechanisms Module Id M28: Translation and Post-translation Modifications in Eukaryotes Keywords Genome, Proteome diversity, post-translational modifications, glycosylation, phosphorylation, methylation Contents 1. Learning Objectives 2. Introduction 3. Purpose of post translational modifications 4. Post translational modifications 4.1. Phosphorylation, the addition of a phosphate group 4.2. Methylation, the addition of a methyl group 4.3. Glycosylation, the addition of sugar groups 4.4. Disulfide bonds, the formation of covalent bonds between 2 cysteine amino acids 4.5. Proteolysis/ Proteolytic Cleavage 4.6. Subunit binding to form a multisubunit protein 4.7. S-nitrosylation 4.8. Lipidation 4.9. Acetylation 4.10. Ubiquitylation 4.11. SUMOlytion 4.12. Vitamin C-Dependent Modifications 4.13. Vitamin K-Dependent Modifications 4.14. Selenoproteins 4.15. Myristoylation 5. Chaperones: Role in PTM and mechanism 6. Role of PTMs in diseases 7. Detecting and Quantifying Post-Translational Modifications 8. -
Protein Targeting Or Protein Sorting
Protein targeting or Protein sorting Refer Page 1068 to 1074 Principles of Biochemistry by Lehninger & Page 663 Baltimore Mol Cell Biology • Protein targeting or Protein sorting is the process of delivery of newly synthesized proteins to their proper cellular destinations • Proteins are sorted to the endoplasmic reticulum (ER), mitochondria, chloroplasts, lysosomes, peroxisomes, and the nucleus by different mechanisms • The process can occur either during protein synthesis or soon after synthesis of proteins by translation at the ribosome. • Most of the integral membrane proteins, secretory proteins and lysosomal proteins are sorted to ER lumen from where these proteins are modified for further sorting • For membrane proteins, targeting leads to insertion of the protein into the lipid bilayer • For secretory/water-soluble proteins, targeting leads to translocation of the entire protein across the membrane into the aqueous interior of the organelle. • Protein destined for cytosol simply remain where they are synthesized • Mitochondrial and chloroplast proteins are first completely synthesized and released from ribosomes. These are then bound by cytosolic chaperone proteins and delivered to receptor on target organelle. • Nuclear proteins such as DNA and RNA polymerases, histones, topoisomerases and proteins that regulate gene expression contain Nuclear localization signal (NLS) which is not removed after the protein is translocated. Unlike ER localization signal sequence which is at N terminal, NLS ca be located almost anywhere along the -
SUMO and Ubiquitin in the Nucleus: Different Functions, Similar Mechanisms?
Downloaded from genesdev.cshlp.org on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW SUMO and ubiquitin in the nucleus: different functions, similar mechanisms? Grace Gill1 Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA The small ubiquitin-related modifier SUMO posttrans- tin, SUMO modification regulates protein localization lationally modifies many proteins with roles in diverse and activity. processes including regulation of transcription, chroma- This review focuses on recent advances in our under- tin structure, and DNA repair. Similar to nonproteolytic standing of SUMO function and regulation, drawing on a roles of ubiquitin, SUMO modification regulates protein limited set of examples relating to gene expression, chro- localization and activity. Some proteins can be modified matin structure, and DNA repair. Comparison of SUMO by SUMO and ubiquitin, but with distinct functional and ubiquitin activities in the nucleus reveals interest- consequences. It is possible that the effects of ubiquiti- ing differences in function and suggests surprising simi- nation and SUMOylation are both largely due to binding larities in mechanism. Thus, for example, modification of proteins bearing specific interaction domains. Both of transcription factors and histones by ubiquitin is gen- modifications are reversible, and in some cases dynamic erally associated with increased gene expression whereas cycles of modification may be required for activity. Stud- modification of transcription factors and histones by ies of SUMO and ubiquitin in the nucleus are yielding SUMO is generally associated with decreased gene ex- new insights into regulation of gene expression, genome pression. In some cases, SUMO and ubiquitin may di- maintenance, and signal transduction. -
The UBE2L3 Ubiquitin Conjugating Enzyme: Interplay with Inflammasome Signalling and Bacterial Ubiquitin Ligases
The UBE2L3 ubiquitin conjugating enzyme: interplay with inflammasome signalling and bacterial ubiquitin ligases Matthew James George Eldridge 2018 Imperial College London Department of Medicine Submitted to Imperial College London for the degree of Doctor of Philosophy 1 Abstract Inflammasome-controlled immune responses such as IL-1β release and pyroptosis play key roles in antimicrobial immunity and are heavily implicated in multiple hereditary autoimmune diseases. Despite extensive knowledge of the mechanisms regulating inflammasome activation, many downstream responses remain poorly understood or uncharacterised. The cysteine protease caspase-1 is the executor of inflammasome responses, therefore identifying and characterising its substrates is vital for better understanding of inflammasome-mediated effector mechanisms. Using unbiased proteomics, the Shenoy grouped identified the ubiquitin conjugating enzyme UBE2L3 as a target of caspase-1. In this work, I have confirmed UBE2L3 as an indirect target of caspase-1 and characterised its role in inflammasomes-mediated immune responses. I show that UBE2L3 functions in the negative regulation of cellular pro-IL-1 via the ubiquitin- proteasome system. Following inflammatory stimuli, UBE2L3 assists in the ubiquitylation and degradation of newly produced pro-IL-1. However, in response to caspase-1 activation, UBE2L3 is itself targeted for degradation by the proteasome in a caspase-1-dependent manner, thereby liberating an additional pool of IL-1 which may be processed and released. UBE2L3 therefore acts a molecular rheostat, conferring caspase-1 an additional level of control over this potent cytokine, ensuring that it is efficiently secreted only in appropriate circumstances. These findings on UBE2L3 have implications for IL-1- driven pathology in hereditary fever syndromes, and autoinflammatory conditions associated with UBE2L3 polymorphisms. -
Celastrol Increases Glucocerebrosidase Activity in Gaucher Disease by Modulating Molecular Chaperones
Celastrol increases glucocerebrosidase activity in Gaucher disease by modulating molecular chaperones Chunzhang Yanga,1, Cody L. Swallowsa, Chao Zhanga, Jie Lua, Hongbin Xiaob, Roscoe O. Bradya,1, and Zhengping Zhuanga,1 aSurgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892-1260; and bInstitute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China Contributed by Roscoe O. Brady, November 19, 2013 (sent for review October 21, 2013) Gaucher disease is caused by mutations in the glucosidase, beta, acid increased the catalytic activity of mutant GCase. Celastrol interfered gene that encodes glucocerebrosidase (GCase). Glucosidase, beta, with the recruitment of Cdc37 to Hsp90 halting the assembly of the acid mutations often cause protein misfolding and quantitative loss requisite chaperone complex. Inhibition of Hsp90 reduced its rec- of GCase. In the present study, we found that celastrol, an herb de- ognition of mutant GCase and therefore limited the proteasomal rivative with known anticancer, anti-inflammatory, and antioxidant degradation of the mutant protein. Additionally, celastrol triggered activity, significantly increased the quantity and catalytic activity of a reorganization of the gene expression pattern of molecular chap- GCase. Celastrol interfered with the establishment of the heat-shock erones such as DnaJ homolog subfamily B members 1 and 9 protein 90/Hsp90 cochaperone Cdc37/Hsp90-Hsp70-organizing pro- (DNAJB1/9), heat shock 70kDa proteins 1A and 1B (HSPA1A/B), tein chaperone complex with mutant GCase and reduced heat-shock and Bcl2-associated athanogene 3 (BAG3). The presence of BAG protein 90-associated protein degradation. In addition, celastrol mod- family molecular chaperone regulator 3 (BAG3) further stabi- ulated the expression of molecular chaperones. -
Loss of Conserved Ubiquitylation Sites in Conserved Proteins During Human Evolution
INTERNATIONAL JOURNAL OF MOleCular meDICine 42: 2203-2212, 2018 Loss of conserved ubiquitylation sites in conserved proteins during human evolution DONGBIN PARK, CHUL JUN GOH, HYEIN KIM, JI SEOK LEE and YOONSOO HAHN Department of Life Science, Chung‑Ang University, Seoul 06974, Republic of Korea Received January 30, 2018; Accepted July 6, 2018 DOI: 10.3892/ijmm.2018.3772 Abstract. Ubiquitylation of lysine residues in proteins serves Introduction a pivotal role in the efficient removal of misfolded or unused proteins and in the control of various regulatory pathways Ubiquitylation, in which the highly conserved 76‑residue poly- by monitoring protein activity that may lead to protein peptide ubiquitin is covalently attached to a lysine residue of degradation. The loss of ubiquitylated lysines may affect substrate proteins, mediates the targeted destruction of ubiq- the ubiquitin‑mediated regulatory network and result in the uitylated proteins by the ubiquitin‑proteasome system (1‑4). emergence of novel phenotypes. The present study analyzed The ubiquitin‑mediated protein degradation pathway serves a mouse ubiquitylation data and orthologous proteins from crucial role in the efficient and specific removal of misfolded 62 mammals to identify 193 conserved ubiquitylation sites from proteins and certain key regulatory proteins (5,6). Ubiquitin 169 proteins that were lost in the Euarchonta lineage leading and other ubiquitin‑like proteins, including autophagy‑related to humans. A total of 8 proteins, including betaine homo- protein 8, Ubiquitin‑like -
Targeting the Ubiquitin System in Glioblastoma', Frontiers in Oncology
Citation for published version: Licchesi, J 2020, 'Targeting the Ubiquitin System in Glioblastoma', Frontiers in Oncology. https://doi.org/10.3389/fonc.2020.574011 DOI: 10.3389/fonc.2020.574011 Publication date: 2020 Document Version Publisher's PDF, also known as Version of record Link to publication University of Bath Alternative formats If you require this document in an alternative format, please contact: [email protected] General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Download date: 24. Sep. 2021 REVIEW published: 25 November 2020 doi: 10.3389/fonc.2020.574011 Targeting the Ubiquitin System in Glioblastoma Nico Scholz 1, Kathreena M. Kurian 2, Florian A. Siebzehnrubl 3 and Julien D. F. Licchesi 1* 1 Department of Biology & Biochemistry, University of Bath, Bath, United Kingdom, 2 Brain Tumour Research Group, Institute of Clinical Neurosciences, University of Bristol, Bristol, United Kingdom, 3 Cardiff University School of Biosciences, European Cancer Stem Cell Research Institute, Cardiff, United Kingdom Glioblastoma is the most common primary brain tumor in adults with poor overall outcome and 5-year survival of less than 5%. Treatment has not changed much in the last decade or so, with surgical resection and radio/chemotherapy being the main options. -
Huang Lab Report
www.beatson.gla.ac.uk/danny_huang (Figure 2A). Upon DNA damage, Ser429 Figure 1 phosphorylation enhanced MDM2 Enzymatic cascade for Ub autoubiquitination and degradation in cells modifications explaining the rapid p53 activation. We also UBIQUITIN SIGNALLING described a strategy for preparation of MDM2 RING domain for structural analyses to enable rapid development MDM2 RING inhibitor. Insights into Deltex ubiquitin ligases Deltex (DTX) family E3s share a highly conserved C-terminal RING domain followed by a Deltex C-terminal domain (DTC). DTXs have been linked Post-translational modification with ubiquitin (Ub) initiated by to developmental processes involving Notch sequential actions of Ub-activating enzyme (E1), Ub-conjugating signalling and histone ubiquitination during DNA enzyme (E2) and Ub ligase (E3) regulates diverse cellular processes, damage repair. However, their functions remain largely unknown. We discovered that DTX E3s including signal transduction, cell cycle progression, apoptosis and harbour dual activities and catalyse both gene transcription. Deregulation in the Ub pathway is often ubiquitination of ADP-ribosylated substrate and ADP-ribosylation of Ub. We showed that DTX’s associated with human pathogenesis, including cancer. Our group C-terminal DTC domain harbour a conserved Group Leader uses structural biology and biochemical approaches to study the pocket that binds both ADP-ribose (ADPr) and NAD+ (Figure 2B). The DTC domains also bind Danny Huang enzymes in the Ub pathway to understand their regulation, levels of p53, the MDM2 mutant was able to poly-ADP-ribosylated substrates in cells. The restrain p53 activity sufficiently for normal Associate Scientist mechanistic functions and mutation-induced deregulation. We proximity of RING and DTC domain enables the growth. -
Airway Inflammation Via Ubch7 Ndfip1 Regulates Itch Ligase Activity
The Journal of Immunology Ndfip1 Regulates Itch Ligase Activity and Airway Inflammation via UbcH7 Mahesh Kathania,* Minghui Zeng,* Viveka Nand Yadav,† Seyed Javad Moghaddam,‡ Baoli Yang,x and K Venuprasad* The ubiquitin-ligating enzyme (E3) Itch plays a crucial role in the regulation of inflammation, and Itch deficiency leads to severe airway inflammation. However, the molecular mechanisms by which Itch function is regulated remain elusive. In this study, we found that nontypeable Haemophilus influenzae induces the association of Itch with Ndfip1. Both Itch2/2 and Ndfip12/2 mice exhibited severe airway inflammation in response to nontypeable Haemophilus influenza, which was associated with elevated expression of proinflammatory cytokines. Ndfip1 enhanced Itch ligase activity and facilitated Itch-mediated Tak1 ubiquitination. Mechanistically, Ndfip1 facilitated recruitment of ubiquitin-conjugating enzyme (E2) UbcH7 to Itch. The N-terminal region of Ndfip1 binds to UbcH7, whereas the PY motif binds to Itch. Hence, Ndfip1 acts as an adaptor for UbcH7 and Itch. Reconstitution of full-length Ndfip1 but not the mutants that fail to interact with either UbcH7 or Itch, restored the defect in Tak1 ubiquitination and inhibited elevated proinflammatory cytokine expression by Ndfip12/2 cells. These results provide new mechanistic insights into how Itch function is regulated during inflammatory signaling, which could be exploited therapeutically in inflammatory diseases. The Journal of Immunology, 2015, 194: 2160–2167. nflammation is essential for host defense against invading valent attachment of Ub to a conserved cysteine residue in the pathogens, such as viruses and bacteria (1). The inflammatory HECT domain, and this Ub is transferred to the target protein (5).