Craig S. McKay
University of Ottawa
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Featured researches published by Craig S. McKay.
Journal of the American Chemical Society | 2011
David C. Kennedy; Craig S. McKay; Marc C.B. Legault; Dana C. Danielson; Jessie A. Blake; Adrian F. Pegoraro; Albert Stolow; Zoltan Mester; John Paul Pezacki
Copper toxicity is a critical issue in the development of copper-based catalysts for copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reactions for applications in living systems. The effects and related toxicity of copper on mammalian cells are dependent on the ligand environment. Copper complexes can be highly toxic, can induce changes in cellular metabolism, and can be rapidly taken up by cells, all of which can affect their ability to function as catalysts for CuAAC in living systems. Herein, we have evaluated the effects of a number of copper complexes that are typically used to catalyze CuAAC reactions on four human cell lines by measuring mitochondrial activity based on the metabolism of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to study toxicity, inductively coupled plasma mass spectrometry to study cellular uptake, and coherent anti-Stokes Raman scattering (CARS) microscopy to study effects on lipid metabolism. We find that ligand environment around copper influences all three parameters. Interestingly, for the Cu(II)-bis-L-histidine complex (Cu(his)(2)), cellular uptake and metabolic changes are observed with no toxicity after 72 h at micromolar concentrations. Furthermore, we show that under conditions where other copper complexes kill human hepatoma cells, Cu(I)-L-histidine is an effective catalyst for CuAAC labeling of live cells following metabolic incorporation of an alkyne-labeled sugar (Ac(4)ManNAl) into glycosylated proteins expressed on the cell surface. This result suggests that Cu(his)(2) or derivatives thereof have potential for in vivo applications where toxicity as well as catalytic activity are critical factors for successful bioconjugation reactions.
Organic and Biomolecular Chemistry | 2012
Craig S. McKay; Mariya Chigrinova; Jessie A. Blake; John Paul Pezacki
Strain-promoted cycloadditions of cyclic nitrones with biaryl-aza-cyclooctynone (BARAC) proceed with rate constants up to 47.3 M(-1) s(-1), this corresponds to a 47-fold rate enhancement relative to reaction of BARAC with benzyl azide and a 14-fold enhancement over previously reported strain promoted alkyne-nitrone cycloadditions (SPANC). Studies of the SPANC reaction using the linear free energy relationship defined by the Hammett equation demonstrated that the cycloaddition reaction has a rho value of 0.25 ± 0.04, indicating that reaction is not sensitive to substituents and thus should have broad applicability.
Proteome Science | 2010
Ragunath Singaravelu; David R. Blais; Craig S. McKay; John Paul Pezacki
BackgroundHepatitis C virus (HCV) poses a growing threat to global health as it often leads to serious liver diseases and is one of the primary causes for liver transplantation. Currently, no vaccines are available to prevent HCV infection and clinical treatments have limited success. Since HCV has a small proteome, it relies on many host cell proteins to complete its life cycle. In this study, we used a non-directed phenyl sulfonate ester probe (PS4≡) to selectively target a broad range of enzyme families that show differential activity during HCV replication in Huh-7 cells.ResultsThe PS4≡ probe successfully targeted 19 active proteins in nine distinct protein families, some that were predominantly labeled in situ compared to the in vitro labeled cell homogenate. Nine proteins revealed altered activity levels during HCV replication. Some candidates identified, such as heat shock 70 kDa protein 8 (or HSP70 cognate), have been shown to influence viral release and abundance of cellular lipid droplets. Other differentially active PS4≡ targets, such as electron transfer flavoprotein alpha, protein disulfide isomerase A5, and nuclear distribution gene C homolog, constitute novel proteins that potentially mediate HCV propagation.ConclusionsThese findings demonstrate the practicality and versatility of non-directed activity-based protein profiling (ABPP) to complement directed methods and accelerate the discovery of altered protein activities associated with pathological states such as HCV replication. Collectively, these results highlight the ability of in situ ABPP approaches to facilitate the identification of enzymes that are either predominantly or exclusively labeled in living cells. Several of these differentially active enzymes represent possible HCV-host interactions that could be targeted for diagnostic or therapeutic purposes.
Trends in Biotechnology | 2012
David R. Blais; Neda Nasheri; Craig S. McKay; Marc C.B. Legault; John Paul Pezacki
Virologists have benefited from large-scale profiling methods to discover new host–virus interactions and to learn about the mechanisms of pathogenesis. One such technique, referred to as activity-based protein profiling (ABPP), uses active site-directed probes to monitor the functional state of enzymes, taking into account post-translational interactions and modifications. ABPP gives insight into the catalytic activity of enzyme families that does not necessarily correlate with protein abundance. ABPP has been used to investigate several viruses and their interactions with their hosts. Differential enzymatic activity induced by viruses has been monitored using ABPP. In this review, we present recent advances and trends involving the use of ABPP methods in understanding host–virus interactions and in identifying novel targets for diagnostic and therapeutic applications.
RSC Advances | 2014
Allison R. Sherratt; Mariya Chigrinova; Craig S. McKay; Louis-Philippe Beaulieu; Yanouchka Rouleau; John Paul Pezacki
An adapted biocompatible version of the Kinugasa reaction, the copper-catalysed alkyne-nitrone cycloaddition followed by rearrangement (CuANCR), was developed for live-cell labelling. CuANCR labelling was demonstrated for both mammalian and bacterial cells. A method for metabolic incorporation of the nitrone group is also described.
ChemBioChem | 2014
Neda Nasheri; Craig S. McKay; Kelly M. Fulton; Susan M. Twine; Megan H. Powdrill; Allison R. Sherratt; John Paul Pezacki
Activity-based protein profiling (ABPP) has emerged as a modern strategy for performing functional proteomics that utilises active-site-directed molecular probes to characterise and assign function to enzyme families within complex proteomes, as well as to discover novel customised enzyme inhibitors. The activities of a variety of enzyme classes have been studied with this technique, including proteases, kinases, phosphatases, hydrolases, and histone deacetylases. 5, b-Lactam-containing compounds, such as penicillin, are a commonly used class of antibiotics against bacterial infections. It has been demonstrated that b-lactams inhibit bacterial growth by targeting the penicillin-binding proteins (PBPs) that are required for bacterial cell wall biosynthesis. b-Lactam-containing activity-based probes, as well as other scaffolds, have been recently used to study bacterial virulence and host responses. Elegant work by Sieber and Staub has shown that b-lactam analogues, both natural and synthetic, can be converted into activity-based probes and applied for comparative in situ profiling of prokaryotic proteomes to identify other enzymatic targets, as well as for determining bacterial sensitivity towards antibiotics. Interestingly, these studies demonstrated that, in addition to PBPs, a broad range of bacterial enzymes that contain a nucleophilic residue in their active pockets can be targeted by the electrophilic blactam ring. 10] Highly similar b-lactone-containing molecules have also been used as activity-based probes; for example, activity-based probes were developed based on Orlistat, the drug that targets the thioesterase domain of human fatty acid synthase (FASN), and were used to study the role of FASN enzyme activity during hepatitis C virus replication and infection. To expand the scope of blactam-containing activity-based probes, and to examine the role of hydrophobicity in directing b-lactam-containing probes
ACS Infectious Diseases | 2018
Geneviève F. Desrochers; Christina Cornacchia; Craig S. McKay; John Paul Pezacki
Protein-protein interactions are integral to host-virus interactions and can contribute significantly to enzyme regulation by changing the localization of both host and viral enzymes within the cell, inducing conformational change relevant to enzyme activity or recruiting other additional proteins to form functional complexes. Identifying the interactors of active enzymes using an activity-based protein profiling probe has allowed us to characterize both normal enzyme activation mechanisms and the manner by which these mechanisms are hijacked and altered by the hepatitis C virus (HCV). Here, we report use of a novel activity-based probe, PIKBPyne, which labels phosphatidylinositol kinases (PIKs) in an activity-based manner, to investigate HCV-dependent changes in protein-protein interactions for PI4KB. Herein, we report the synthesis of new variations on PIKBPyne, compare their ability to label the interacting partners of PI4KB, and demonstrate the utility of our approach in characterizing virus-mediated changes to host function.
Chemical Communications | 2010
Craig S. McKay; Joseph Moran; John Paul Pezacki
Chemical Communications | 2011
Craig S. McKay; Jessie A. Blake; Jenny Cheng; Dana C. Danielson; John Paul Pezacki
Tetrahedron Letters | 2009
Craig S. McKay; David C. Kennedy; John Paul Pezacki