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Engineered Glucose to Generate a Spectroscopic Probe for Studying Carbohydrate Biology

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Biochemical Roles of Eukaryotic Cell Surface Macromolecules

Part of the book series: Advances in Experimental Medicine and Biology ((volume 749))

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Abstract

Metabolic carbohydrate engineering by exogenously added monosaccharide supplement is a technique of importance for studying physiological role of various glycans. Additionally, it also has the potential of developing new drug molecules for specific targeting. Lack of a spectroscopic reporting moiety in carbohydrates makes understanding their biochemical and physiological role very difficult. Towards this goal, we have modified glucose, with a propargyl group, wherein an azido coumarinyl profluorophore has been linked by “click chemistry.” Here, we demonstrate the uptake and incorporation of this modified monosaccharide into bacteria, yeast and mammalian cells. We show that modification at C-2 (carbon numbered 2, according to IUPAC) position is tolerated best, and uptake is only slightly lower compared to glucose. In the presence of C-2-modified glucose, growth kinetics and cellular viability were also minimally affected in all the cell types used. Fluorescence spectroscopy of the labeled biomolecule and fluorescence imaging of the cells demonstrate that C-2-modified glucose is metabolically incorporated not only in the cell membrane but also accumulates in the nucleus. Such a fluorophore, incorporated into biomolecules, can be used as a tool to understand their structure–function relationship. Here, we show that the incorporation of such a fluorophore in a carbohydrate moiety may enable studying the physiological and biochemical processes associated with membranes.

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Acknowledgments

SH thanks Director, NCL for financial support for LC-MS facility. The authors thank Dr. Sathyanarayana Gummadi for critical reading of the manuscript and encouragement. GKA thanks IIT Madras for the financial support. AT thanks the fellowship from CSIR-New Delhi.

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Authors and Affiliations

  1. Division of Organic Chemistry, Combi Chem—Bio Resource Center, National Chemical Laboratory, PUNE, 411 008, India

    Ashish Tripathi & Srinivas Hotha

  2. Department of Biotechnology, Indian Institute of Technology Madras, Chennai, 600 036, India

    Vibha Singh, K. G. Aishwarya & Gopala Krishna Aradhyam

Authors
  1. Ashish Tripathi
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  2. Vibha Singh
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  3. K. G. Aishwarya
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  4. Gopala Krishna Aradhyam
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  5. Srinivas Hotha
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Corresponding authors

Correspondence to Gopala Krishna Aradhyam or Srinivas Hotha .

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Editors and Affiliations

  1. Riverside Transit Campus, School of Biological Sciences, Central University of Kerala, Kasargod, 671328, Kerala, India

    Perumana R. Sudhakaran

  2. National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067, Delhi, India

    Avadhesha Surolia

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Tripathi, A., Singh, V., Aishwarya, K.G., Aradhyam, G.K., Hotha, S. (2012). Engineered Glucose to Generate a Spectroscopic Probe for Studying Carbohydrate Biology. In: Sudhakaran, P., Surolia, A. (eds) Biochemical Roles of Eukaryotic Cell Surface Macromolecules. Advances in Experimental Medicine and Biology, vol 749. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-3381-1_21

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