Methyl Arginine Processing Enzymes: Small Molecule Modulation, Biological Mechanisms and Therapeutic Applications
Lead Research Organisation:
University College London
Department Name: Chemistry
Abstract
Methylation of arginine residues in proteins is increasingly recognised as an important post-translational modification (PTM). The reaction is catalysed by a family of protein arginine methyltransferases (PRMTs) that methylate the guanidine nitrogens of arginine. Two broad families of PRMTs have been described / type I that generates asymmetric dimethylarginine (ADMA) and type II that generates symmetric dimethylarginine (SDMA). Both types of PRMTs can also generate monomethyl arginine (LNMMA), probably as an intermediate en route to dimethylation. Recently there has been increased interest in arginine methylation for three reasons: first because hydrolysis of proteins containing methylarginines leads to the generation of free ADMA and L-NMMA, both of which can inhibit nitric oxide synthase (NOS) enzymes and thereby influence intracellular signalling. Secondly because it has been found that an arginine deiminase can metabolise methylarginine residues in proteins to citrulline and therefore the modification is reversible and may be analogous to protein phosphorylation. Thirdly certain isoforms of the enzymes which process methylarginines i.e. dimethylarginine dimethylaminohydrolase (DDAH); arginine deiminase (ADI) and peptidyl arginine deiminase have been implicated in basic biochemical pathways of pathogenic bacteria. Thus the functional significance of the pathways related to arginine methylation and demethylation appears considerable and may be implicated in fundamental cellular processes such as cell cycle control as well as being implicated in disease processes ranging from cancer to coronary heart disease to bacterial infection. Building on our extensive experience in this area we propose to:The applicants propose to carry out an overarching programme of work to study these enzymes. In this we will integrate activities in chemistry, biochemistry, biophysics, NMR, crystallography, pharmacology and experimental medicine to achieve the following.(1) To extend our recent exciting findings which have identified novel small molecule inhibitors / modulators of these enzymes / such molecules will have potential therapeutic value in a variety of disease states, in particular bacterial infection and cancer. To date our activities have identified novel small molecule structures which selectively inhibit either the human (published) or bacterial form (unpublished) of these enzymes. We have also identified the first small molecule inhibitors of the bacterial enzyme ADI (unpublished). (2) We propose to carefully delineate the nature of the interaction of a variety of new small molecule entities with these methylarginine processing enzymes using Biophysical techniques (Ladbury), Crystallography (McDonald) and NMR. NMR studies on DDAH (Driscoll) and ITC (Ladbury) studies on DDAH have already helped determine the relative positions of binding of natural and non-natural small molecules. Moreover two of the team (McDonald / Vallance) solved the first structure of the bacterial form of DDAH and currently have crystals of the first small-molecule inhibitors bound to DDAH.(3) We propose to evaluate the relevance of these novel small molecule-protein interactions in vivo. We will evaluate their effectiveness as anti-bacterial agents, their ability to enter cells (using appropriately labelled entities, and a variety of microscopy techniques) and the ability to modulate nitric oxide levels, for example in endothelial cells.
Organisations
Publications
Adams AL
(2013)
Cysteine promoted C-terminal hydrazinolysis of native peptides and proteins.
in Angewandte Chemie (International ed. in English)
Baker JR
(2007)
Protein-small molecule interactions in neocarzinostatin, the prototypical enediyne chromoprotein antibiotic.
in Chembiochem : a European journal of chemical biology
Baker JR
(2010)
An efficient asymmetric synthesis of the potent beta-blocker ICI-118,551 allows the determination of enantiomer dependency on biological activity.
in Chemical communications (Cambridge, England)
Cawley MJ
(2008)
Development of a practical Buchwald-Hartwig amine arylation protocol using a conveniently prepared (NHC)Pd(R-allyl)Cl catalyst.
in Organic & biomolecular chemistry
Chudasama V
(2010)
Hydroacylation of alpha,beta-unsaturated esters via aerobic C-H activation.
in Nature chemistry
Esposito O
(2007)
Synthesis and reactivity of alkyl-palladium N-heterocyclic carbene complexes.
in Chemical communications (Cambridge, England)
Esposito O
(2007)
Synthesis and reactivity of alkyl-palladium N-heterocyclic carbene complexes.
in Chemical communications (Cambridge, England)
Gois PM
(2007)
Tuning the reactivity of dirhodium(II) complexes with axial N-heterocyclic carbene ligands: the arylation of aldehydes.
in Angewandte Chemie (International ed. in English)
Gois PM
(2007)
Tuning the reactivity of dirhodium(II) complexes with axial N-heterocyclic carbene ligands: the arylation of aldehydes.
in Angewandte Chemie (International ed. in English)
Knight E
(2016)
Synthesis of novel and potent vorapaxar analogues.
in Organic & biomolecular chemistry
Moody P
(2012)
Bromomaleimide-linked bioconjugates are cleavable in mammalian cells.
in Chembiochem : a European journal of chemical biology
Nathani R
(2012)
Bioconjugation of green fluorescent protein via an unexpectedly stable cyclic sulfonium intermediate.
in Chembiochem : a European journal of chemical biology
Nathani RI
(2013)
A novel approach to the site-selective dual labelling of a protein via chemoselective cysteine modification.
in Chemical science
Robinson E
(2016)
Identification of an active metabolite of PAR-1 antagonist RWJ-58259 and synthesis of analogues to enhance its metabolic stability.
in Organic & biomolecular chemistry
Ryan CP
(2011)
Tunable reagents for multi-functional bioconjugation: reversible or permanent chemical modification of proteins and peptides by control of maleimide hydrolysis.
in Chemical communications (Cambridge, England)
Smith ME
(2010)
Protein modification, bioconjugation, and disulfide bridging using bromomaleimides.
in Journal of the American Chemical Society
Wang Z
(2014)
Pharmacological inhibition of DDAH1 improves survival, haemodynamics and organ function in experimental septic shock.
in The Biochemical journal
Description | As part of this award we discovered a number of new approaches to synthetic modification of proteins which has broad implications for chemical biology. Many of these have now been extended into published works and patents. |
Exploitation Route | Some of these findings have been further progressed though a variety of research grants and PhD studentships. We also have extended the findings into patents and through a variety of PoC and other translational funds have now developed a technology platform. |
Sectors | Chemicals Pharmaceuticals and Medical Biotechnology |
URL | https://www.ucl.ac.uk/caddick-group |
Description | The work has led to a number of patents and these have now led to license agreements - patents are now being granted and technology is advancing through a series of PoC awards and investments. |
First Year Of Impact | 2010 |
Sector | Chemicals,Pharmaceuticals and Medical Biotechnology |
Impact Types | Societal Economic |
Description | BBSRC Grouped |
Amount | £123,000 (GBP) |
Funding ID | BB/FOF/313 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2010 |
End | 03/2011 |
Description | BBSRC Grouped |
Amount | £123,000 (GBP) |
Funding ID | BB/FOF/313 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2010 |
End | 03/2011 |
Title | Functionalisation of Solid Substrates |
Description | The present invention relates to a product comprising a solid substrate and a moiety of formula (I) linked thereto: wherein X, X? and R are as defined herein. The product is useful for immobilising target molecules such as molecules of biochemical interest to solid substrates for numerous applications, such as affinity chromatography, ELISA, biotechnological assay techniques and solid phase peptide synthesis. |
IP Reference | US20120190579 |
Protection | Patent application published |
Year Protection Granted | 2010 |
Licensed | No |
Impact | None |
Title | Reversible Covalent Linkage of Functional Molecules |
Description | Abstract: The present invention relates to the use of a compound containing a moiety of formula (I) as a reagent for linking a compound of formula R1-H which comprises a first functional moiety of formula F1 to a second functional moiety of formula F2 wherein X, X?, Y, R1, F1 and F2 are as defined herein. The present invention also provides related processes and products. The present invention is useful for creating functional conjugate compounds, and specifically conjugates in which at least one of the constituent molecules carries a thiol group. |
IP Reference | US20120190124 |
Protection | Patent application published |
Year Protection Granted | 2010 |
Licensed | Commercial In Confidence |
Impact | Patent has been granted and further work has carried on and been funded and led to new patents and a spinout company - Thiologics. |
Title | Thiol Protecting Group |
Description | Abstract: The present invention relates to the use of a compound comprising a moiety of formula (I) as a reagent for protecting a thiol group in a thiol compound wherein X, X? and Y are as defined herein. The protecting group methodology of the present invention allows straightforward and selective protection of thiol groups. Cleaving of the thiol group to regenerate the thiol functional group is also facile and controllable. The present invention further provides an analogous protecting group methodology directed to protection of disulfide groups. |
IP Reference | US20120190814 |
Protection | Patent application published |
Year Protection Granted | 2010 |
Licensed | No |
Impact | None |