Senior Research Scientist
Nigel Richards

Education:

BSc (Hons) in Chemistry, Imperial College, University of London, UK (1980)
PhD in Organic Synthesis, University of Cambridge, UK (1983)
Harkness Fellow, Computational Chemistry, Columbia University, USA (1983-1985)

Research Summary:

My research focuses on understanding the structure, mechanism, evolution and applications of enzymes. Current projects include:

Computational investigations of residue networks that mediate energy transfer in enzymes
Structural and mechanistic characterization of enzymes that mediate C-nucleoside biosynthesis
Biocatalytic routes to novel C-nucleosides with potential use as anti-viral agents
Discovery and characterization of asparagine synthetase inhibitors as anti-cancer agents
Computational simulations of DNA and RNA containing modified nucleotides from an Artificially Expanded Genetic Information System (AEGIS)
Characterization and engineering of psychrophilic DNA polymerases and DNA ligases

Awards

Fellow, American Association for the Advancement of Science, 2010
Fellow, Royal Society of Chemistry, 2016
Honorary Fellow, Indian Society of Chemists & Biologists, 2020
Fellow, Learned Society of Wales, 2021

Recent Publications

The chemistry of formycin biosynthesis
Richards NGJ, Naismith JN
Front. Chem. Biol., Frontiers Media SA (2024) 11 July: 3, 1428646. DOI: 10.3389/fchbi.2024.1428646
<Abstract>

Remarkable progress has been made to elucidate the structural and mechanistic enzymology of the biosynthetic pathways that give rise to naturally occurring C-nucleosides. These compounds are generally cytotoxic and exhibit interesting antiviral, antibiotic and anti-parasitic activity. Here we review current knowledge concerning formycin biosynthesis and highlight deficiencies in our understanding of key chemical transformations in the pathway.

Arginine kinase activates arginine for phosphorylation by pyramidalization and polarization
Falcioni F, Molt RW Jr., Jin Y, Waltho JP, Hay S, Richards, NGJ, Blackburn GM
ACS Catal, ACS (2024) 16 April: 14(9), 6650-6658. DOI: 10.1021/acscatal.4c00380
<Abstract>

Arginine phosphorylation plays numerous roles throughout biology. Arginine kinase (AK) catalyzes the delivery of an anionic phosphoryl group (PO₃^–) from ATP to a planar, trigonal nitrogen in a guanidinium cation. Density functional theory (DFT) calculations have yielded a model of the transition state (TS) for the AK-catalyzed reaction. They reveal a network of over 50 hydrogen bonds that delivers unprecedented pyramidalization and out-of-plane polarization of the arginine guanidinium nitrogen (Nη2) and aligns the electron density on Nη2 with the scissile P–O bond, leading to in-line phosphoryl transfer via an associative mechanism. In the reverse reaction, the hydrogen-bonding network enforces the conformational distortion of a bound phosphoarginine substrate to increase the basicity of Nη2. This enables Nη2 protonation, which triggers PO₃^– migration to generate ATP. This polarization–pyramidalization of nitrogen in the arginine side chain is likely a general phenomenon that is exploited by many classes of enzymes mediating the post-translational modification of arginine.

Reactivity and mechanism in chemical and synthetic biology
Richards, NGJ, Bearne SL, Goto Y, Parker EJ
Phil. Trans. R. Soc. B Biol. Sci., The Royal Society Publishing (2023) 27 February: 378(1871), 20220023. DOI: 10.1098/rstb.2022.0023
<Abstract>

Physical organic chemistry and mechanistic thinking provide a strong intellectual framework for understanding the chemical logic of evolvable informational macromolecules and metabolic transformations in living organisms. These concepts have also led to numerous successes in designing and applying tools to delineate biological function in health and disease, chemical ecology and possible alternative chemistries employed by extraterrestrial life. A symposium at the 2020 Pacifichem meeting was scheduled in December 2020 to discuss designing and exploiting expanded genetic alphabets, methods to understand the biosynthesis of natural products and re-engineering primary metabolism in bacteria. The COVID-19 pandemic led to postponement of in-person discussions, with the symposium eventually being held on 20–21 December 2021 as an online event. This issue is a written record of work presented on biosynthetic pathways and enzyme catalysis, engineering microorganisms with new metabolic capabilities, and the synthesis of non-canonical, nucleobases for medical applications and for studies of alternate chemistries for living organisms. The variety of opinion pieces, reviews and original research articles provide a starting point for innovations that clarify how complex biological systems emerge from the rules of chemical reactivity and mechanism.

Experimental and computational snapshots of C-C bond formation in a C-nucleoside synthase
Li W, Girt GC, Radadiya A, Stewart JJP, Richards NGJ, Naismith JN
Open Biology, The Royal Society Publishing (2023) 11 January: 13(1), 220287. DOI: 10.1098/rsob.220287
<Abstract>

The biosynthetic enzyme, ForT, catalyses the formation of a C-C bond between 4-amino-1H-pyrazoledicarboxylic acid and MgPRPP to produce a C-nucleoside precursor of formycin A. The transformation catalysed by ForT is of chemical interest because it is one of only a few examples in which C-C bond formation takes place via an electrophilic substitution of a small, aromatic heterocycle. In addition, ForT is capable of discriminating between the aminopyrazoledicarboxylic acid and an analogue in which the amine is replaced by a hydroxyl group; a remarkable feat given the steric and electronic similarities of the two molecules. Here we report biophysical measurements, structural biology and quantum chemical calculations that provide a detailed molecular picture of ForT-catalysed C-C bond formation and the conformational changes that are coupled to catalysis. Our findings set the scene for employing engineered ForT variants in the biocatalytic production of novel, anti-viral C-nucleoside and C-nucleotide analogues.

Development of triazole-conjugated dihydropyrimidinone (DHPM) derivatives as potential P-glycoprotein inhibitors
Bijani S, Shaikh F, Mirza D, Siu SWI, Jain N, Ferreira RJ, dos Santos DJVA, Rawal R, Richards NGJ, Shah A, Radadiya A
ACS Omega, ACS (2022) 17 May: 7(19), 16278-16287. DOI: 10.1021/acsomega.1c05839
<Abstract>

P-glycoprotein (Pgp), an ATP binding cassette (ABC) transporter, is an ATP-dependent efflux pump responsible for cancer multidrug resistance. As part of efforts to identify human Pgp (hPgp) inhibitors, we prepared a series of novel triazole-conjugated dihydropyrimidinones using a synthetic approach that is well suited for obtaining compound libraries. Several of these dihydropyrimidinone derivatives modulate human P-glycoprotein (hPgp) activity with low micromolar EC50 values. Molecular docking studies suggest that these compounds bind to the M-site of the transporter.

Functional characterization of two PLP-dependent enzymes involved in capsular polysaccharide biosynthesis from Campylobacter jejuni
Riegert AS, Narindoshvili T, Coricello A, Richards NGJ, Raushel FM
Biochemistry, ACS (2021) 21 September: 60(19), 2836-2843. DOI: 10.1021/acs.biochem.1c00439
<Abstract>

Campylobacter jejuni is a Gram-negative, pathogenic bacterium that causes campylobacteriosis, a form of gastroenteritis. C. jejuni is the most frequent cause of food-borne illness in the world, surpassing Salmonella and E. coli. Coating the surface of C. jejuni is a layer of sugar molecules known as the capsular polysaccharide that, in C. jejuni NCTC 11168, is composed of a repeating unit of d-glycero-l-gluco-heptose, d-glucuronic acid, d-N-acetyl-galactosamine, and d-ribose. The d-glucuronic acid moiety is further amidated with either serinol or ethanolamine. It is unknown how these modifications are synthesized and attached to the polysaccharide. Here, we report the catalytic activities of two previously uncharacterized, pyridoxal phosphate (PLP)-dependent enzymes, Cj1436 and Cj1437, from C. jejuni NCTC 11168. Using a combination of mass spectrometry and nuclear magnetic resonance, we determined that Cj1436 catalyzes the decarboxylation of l-serine phosphate to ethanolamine phosphate. Cj1437 was shown to catalyze the transamination of dihydroxyacetone phosphate to (S)-serinol phosphate in the presence of l-glutamate. The probable routes to the ultimate formation of the glucuronamide substructures in the capsular polysaccharides of C. jejuni are discussed.

Building better polymerases: Engineering the replication of expanded genetic alphabets
Ouaray, Z., Benner, S. A., Georgiadis, M. M., Richards, N. G. J.
J. Biol. Chem., ASBMB (2020) 11 December: 295(50):17046-17059, DOI:10.1074/jbc.REV120.013745
<Abstract>

DNA polymerases are today used throughout scientific research, biotechnology, and medicine, in part for their ability to interact with unnatural forms of DNA created by synthetic biologists. Here especially, natural DNA polymerases often do not have the "performance specifications" needed for transformative technologies. This creates a need for science-guided rational (or semi-rational) engineering to identify variants that replicate unnatural base pairs (UBPs), unnatural backbones, tags, or other evolutionarily novel features of unnatural DNA. In this review, we provide a brief overview of the chemistry and properties of replicative DNA polymerases and their evolved variants, focusing on the Klenow fragment of Taq DNA polymerase (Klentaq). We describe comparative structural, enzymatic, and molecular dynamics studies of WT and Klentaq variants, complexed with natural or noncanonical substrates. Combining these methods provides insight into how specific amino acid substitutions distant from the active site in a Klentaq DNA polymerase variant (ZP Klentaq) contribute to its ability to replicate UBPs with improved efficiency compared with Klentaq. This approach can therefore serve to guide any future rational engineering of replicative DNA polymerases.

Improving the treatment of acute lymphoblastic leukemia
Radadiya A, Zhu W, Coricello A, Alcaro S, Richards NGJ
Biochemistry, ACS (2020) 8 September: 59(35), 3193-3200. DOI: 10.1021/acs.biochem.0c00354
<Abstract>

l-Asparaginase (EC 3.5.1.1) was first used as a component of combination drug therapies to treat acute lymphoblastic leukemia (ALL), a cancer of the blood and bone marrow, almost 50 years ago. Administering this enzyme to reduce asparagine levels in the blood is a cornerstone of modern clinical protocols for ALL; indeed, this remains the only successful example of a therapy targeted against a specific metabolic weakness in any form of cancer. Three problems, however, constrain the clinical use of l-asparaginase. First, a type II bacterial variant of l-asparaginase is administered to patients, the majority of whom are children, which produces an immune response thereby limiting the time over which the enzyme can be tolerated. Second, l-asparaginase is subject to proteolytic degradation in the blood. Third, toxic side effects are observed, which may be correlated with the l-glutaminase activity of the enzyme. This Perspective will outline how asparagine depletion negatively impacts the growth of leukemic blasts, discuss the structure and mechanism of l-asparaginase, and briefly describe the clinical use of chemically modified forms of clinically useful l-asparaginases, such as Asparlas, which was recently given FDA approval for use in children (babies to young adults) as part of multidrug treatments for ALL. Finally, we review ongoing efforts to engineer l-asparaginase variants with improved therapeutic properties and briefly detail emerging, alternate strategies for the treatment of forms of ALL that are resistant to asparagine depletion.

Uncovering the chemistry of C-C bond formation in C-nucleoside biosynthesis: Crystal structure of a C-glycoside synthase/PRPP complex
Gao S, Radadiya A, Li W, Liu H, Zhu W, de Crecy-Lagard V, Richards NGJ, Naismith JN
Chem. Comm., Royal Society of Chemistry (2020) 14 July: 56(55), 7617-7620. DOI: 10.1039/d0cc02834g
<Abstract>

The enzyme ForT catalyzes C–C bond formation between 5'-phosphoribosyl-1'-pyrophosphate (PRPP) and 4-amino-1H-pyrazole-3,5-dicarboxylate to make a key intermediate in the biosynthesis of formycin A 5'-phosphate by Streptomyces kaniharaensis. We report the 2.5 Å resolution structure of the ForT/PRPP complex and locate active site residues critical for PRPP recognition and catalysis.

Tautomeric equilibria of nucleobases in the hachimoji expanded genetic alphabet
Eberlein, L., Beierlein, F.R., van Eikema Hommes, N.J.R., Radadiya, A., Heil, J., Benner, S.A., Clark, T., Kast, S.M., and Richards, N.G.J.
J. Chem. Theory Comput., ACS (2020) 14 Apr: 16(4):2766-2777, DOI:10.1021/acs.jctc.9b01079
<Abstract>

Evolution has yielded biopolymers that are constructed from exactly four building blocks and are able to support Darwinian evolution. Synthetic biology aims to extend this alphabet, and we recently showed that 8-letter (hachimoji) DNA can support rule-based information encoding. One source of replicative error in non-natural DNA-like systems, however, is the occurrence of alternative tautomeric forms, which pair differently. Unfortunately, little is known about how structural modifications impact free-energy differences between tautomers of the non-natural nucleobases used in the hachimoji expanded genetic alphabet. Determining experimental tautomer ratios is technically difficult, and so, strategies for improving hachimoji DNA replication efficiency will benefit from accurate computational predictions of equilibrium tautomeric ratios. We now report that high-level quantum-chemical calculations in aqueous solution by the embedded cluster reference interaction site model, benchmarked against free-energy molecular simulations for solvation thermodynamics, provide useful quantitative information on the tautomer ratios of both Watson-Crick and hachimoji nucleobases. In agreement with previous computational studies, all four Watson-Crick nucleobases adopt essentially only one tautomer in water. This is not the case, however, for non-natural nucleobases and their analogues. For example, although the enols of isoguanine and a series of related purines are not populated in water, these heterocycles possess N1-H and N3-H keto tautomers that are similar in energy, thereby adversely impacting accurate nucleobase pairing. These robust computational strategies offer a firm basis for improving experimental measurements of tautomeric ratios, which are currently limited to studying molecules that exist only as two tautomers in solution.

(View publication page for Nigel Richards)

Senior Research Scientist
Nigel Richards

Education:

Research Summary:

Awards

Recent Publications

Photo

Interests

Contact

Senior Research Scientist Nigel Richards

Education:

Research Summary:

Awards

Recent Publications

Photo

Interests

Contact

Senior Research Scientist
Nigel Richards