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Glycoengineering of Esterase Activity through Metabolic Flux-Based Modulation of Sialic Acid.
Mathew, Mohit P; Tan, Elaine; Labonte, Jason W; Shah, Shivam; Saeui, Christopher T; Liu, Lingshu; Bhattacharya, Rahul; Bovonratwet, Patawut; Gray, Jeffrey J; Yarema, Kevin J.
Afiliação
  • Mathew MP; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Tan E; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Labonte JW; Department of Chemical and Biochemical Engineering, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Shah S; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Saeui CT; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Liu L; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Bhattacharya R; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Bovonratwet P; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Gray JJ; Department of Chemical and Biochemical Engineering, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
  • Yarema KJ; Department of Biomedical Engineering and, Translational Tissue Engineering Center, The Johns Hopkins University, 5029 Robert H. & Clarice Smith Building, 400 North Broadway, Baltimore, MD, 21231, USA.
Chembiochem ; 18(13): 1204-1215, 2017 07 04.
Article em En | MEDLINE | ID: mdl-28218815
This report describes the metabolic glycoengineering (MGE) of intracellular esterase activity in human colon cancer (LS174T) and Chinese hamster ovary (CHO) cells. In silico analysis of carboxylesterases CES1 and CES2 suggested that these enzymes are modified with sialylated N-glycans, which are proposed to stabilize the active multimeric forms of these enzymes. This premise was supported by treating cells with butanolylated ManNAc to increase sialylation, which in turn increased esterase activity. By contrast, hexosamine analogues not targeted to sialic acid biosynthesis (e.g., butanoylated GlcNAc or GalNAc) had minimal impact. Measurement of mRNA and protein confirmed that esterase activity was controlled through glycosylation and not through transcription or translation. Azide-modified ManNAc analogues widely used in MGE also enhanced esterase activity and provided a way to enrich targeted glycoengineered proteins (such as CES2), thereby providing unambiguous evidence that the compounds were converted to sialosides and installed into the glycan structures of esterases as intended. Overall, this study provides a pioneering example of the modulation of intracellular enzyme activity through MGE, which expands the value of this technology from its current status as a labeling strategy and modulator of cell surface biological events.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Ácidos Siálicos / Hidrolases de Éster Carboxílico / Processamento de Proteína Pós-Traducional / Carboxilesterase / Células Epiteliais / Engenharia Metabólica Limite: Animals / Humans Idioma: En Revista: Chembiochem Assunto da revista: BIOQUIMICA Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos
Texto completo
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Ácidos Siálicos / Hidrolases de Éster Carboxílico / Processamento de Proteína Pós-Traducional / Carboxilesterase / Células Epiteliais / Engenharia Metabólica Limite: Animals / Humans Idioma: En Revista: Chembiochem Assunto da revista: BIOQUIMICA Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos