A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts

PLoS Comput Biol. 2016 Apr 7;12(4):e1004844. doi: 10.1371/journal.pcbi.1004844. eCollection 2016 Apr.

Abstract

O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. Software for generating glycans from the enzyme activities is presented, and is also available online. The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. Simple graph-theoretic measures were used to characterise the resulting reaction networks. From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Computational Biology
  • Computer Simulation
  • Cricetulus
  • Gene Knockout Techniques
  • Genetic Engineering
  • Glycosylation
  • Glycosyltransferases / deficiency
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism
  • Humans
  • Knowledge Bases*
  • Metabolic Networks and Pathways / genetics
  • Models, Biological
  • Mucins / chemistry
  • Mucins / metabolism*
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism
  • Protein Processing, Post-Translational*
  • Terminology as Topic

Substances

  • Mucins
  • Polysaccharides
  • Glycosyltransferases

Grants and funding

This work was part supported by an EU Initial Training Network, Project No. 608381 - Training in Neurodegeneration, Therapeutics Intervention and Neurorepair (TINTIN) awarded to GPD. URL: http://ec.europa.eu/research/fp7/index_en.cfm; and Science Foundation Ireland Grant No. SFI-13/SP SSPC/I2893 URL: http://www.sfi.ie. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.