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Metabolic selection of glycosylation defects in human cells
K J Yarema1, S Goon, C R Bertozzi
1Department of Chemistry, University of California, Berkeley, CA 94720, USA.
Abstract:
Changes in glycosylation are often associated with disease progression, but the genetic and metabolic basis of these events is rarely understood in detail at a molecular level. We describe a metabolism-based approach to the selection of mutants in glycoconjugate biosynthesis that provides insight into regulatory mechanisms for oligosaccharide expression and metabolic flux. Unnatural intermediates are used to challenge a specific pathway, and cell surface expression of their metabolic products provides a readout of flux in that pathway and a basis for selecting genetic mutants. The approach was applied to the sialic acid metabolic pathway in human cells, yielding novel mutants with phenotypes related to the inborn metabolic defect sialuria and metastatic tumor cells.
Insights
Researchers developed a novel metabolism-based method to study glycoconjugate biosynthesis. This approach identifies genetic mutants by tracking unnatural intermediates, offering insights into disease-related metabolic pathways.
Area of Science:
- Biochemistry
- Genetics
- Cell Biology
Background:
- Glycosylation changes are linked to disease, but their molecular basis is often unclear.
- Understanding the genetic and metabolic control of oligosaccharide expression is crucial.
Purpose of the Study:
- To develop a metabolism-based strategy for selecting mutants in glycoconjugate biosynthesis.
- To gain insights into regulatory mechanisms of oligosaccharide expression and metabolic flux.
- To apply this approach to the sialic acid pathway in human cells.
Main Methods:
- Utilizing unnatural metabolic intermediates to probe specific biosynthetic pathways.
- Measuring cell surface expression of metabolic products as a readout of pathway flux.
- Selecting genetic mutants based on altered metabolic product expression.
Main Results:
- Successfully applied the method to the sialic acid metabolic pathway in human cells.
- Identified novel mutants with phenotypes relevant to sialuria and metastatic cancer.
- Demonstrated the utility of unnatural intermediates for pathway analysis and mutant selection.
Conclusions:
- The described metabolism-based approach provides a powerful tool for dissecting complex metabolic pathways.
- This method offers new avenues for understanding diseases associated with altered glycosylation.
- Novel mutants generated can serve as models for studying sialuria and cancer metastasis.