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  • Periodicals (Báo – Tạp chí)

  • Mammal ian Cel l Prol iferation Requires Noncatalytic Functions of O-GlcNAc Transferase

    • Authors : Levine, Zebulon G. (2020)

  • Developed approaches to replace endogenous OGT with separation-of-function variants to investigate the importance of OGT’s enzymatic activities for cell viability. Using genetic complementation, we found that OGT’s glycosyltransferase function is required for cell growth but its protease function is dispensable. We next used complementation to construct a cell line with degron-tagged wild-type OGT. When OGT was degraded to very low levels, cells stopped proliferating b...
  • Journal article

  • Co-Occurrence of Enzyme Domains Guides the Discovery of an Oxazolone Synthetase

    • Authors : Rond, Tristan de; Asay, Julia E.; Moore, Bradley S. (2020)

  • The development of a new genome mining approach for the targeted discovery of novel biochemical transformations through the analysis of co-occurring enzyme domains (CO-ED) in a single protein. CO-ED was designed to identify unannotated multifunctional enzymes for functional characterization and discovery based on the premise that linked enzyme domains have evolved to function collaboratively. Guided by CO-ED, we targeted an unannotated predicted ThiF-nitroreductase di-domai...
  • Periodicals (Báo – Tạp chí)

  • Generation of SARS-CoV-2 S1 spike glycoprotein putative antigenic epitopes in vitro by intracellular aminopeptidases

    • Authors : Stamatakis, George; Samiotaki, Martina; Mpakali, Anastasia (2020)

  • Presentation of antigenic peptides by MHCI is central to cellular immune responses against viral pathogens. While adaptive immune responses versus SARS-CoV-2 can be of critical importance to both recovery and vaccine efficacy, how protein antigens from this pathogen are processed to generate antigenic peptides is largely unknown. Here, we analyzed the proteolytic processing of overlapping precursor peptides spanning the entire sequence of the S1 spike glycoprotein...
  • Journal article

  • A giant leap in sequence space reveals the intracellular complexities of evolving a new function

    • Authors : Hall, Kelsi R; Robins, Katherine J; Rich, Michelle H (2020)

  • Selection for a promiscuous enzyme activity provides substantial opportunity for competition between endogenous and new substrates to influence the evolutionary trajectory, an aspect that has generally been overlooked in laboratory directed evolution studies. We evolved the Escherichia coli nitro/quinone reductase NfsA to detoxify chloramphenicol by randomising eight active site residues simultaneously and interrogating ∼250,000,000 reconfigured NfsA variants. Analysis of every possi...
  • Journal article

  • Naturally occurring cobalamin (B12) analogs can function as cofactors for human methylmalonyl-CoA mutase

    • Authors : Sokolovskaya, Olga M; Plessl, Tanja; Bailey, Henry (2020)

  • Cobalamin, commonly known as vitamin B12, is an essential micronutrient for humans because of its role as an enzyme cofactor. Cobalamin is one of over a dozen structurally related compounds – cobamides – that are found in food and are produced by microorganisms in the human gut. Very little is known about how different cobamides affect B12-dependent metabolism in human cells. Here, we test in vitro how diverse cobamide cofactors affect...