Genomic analysis of biochemical function.

Area of Science:

• Proteomics
• Molecular Biology
• Biochemistry

Background:

• Linking genes to their biochemical activities is crucial for understanding cellular functions.
• Advances in mass spectrometry and affinity purification have improved this linkage.
• Genomic protein arrays offer a high-throughput method for proteome-wide analysis.

Purpose of the Study:

• To establish a robust method for linking individual biochemical activities to their cognate genes.
• To enable the analysis of the entire proteome for biochemical activities.
• To identify genes responsible for specific protein functions through deconvolution.

Main Methods:

• Construction of a yeast genomic protein array by fusing open reading frames to glutathione-S-transferase (GST).
• Purification and biochemical assay of pools of GST-open-reading-frame proteins.
• Analysis of the entire proteome for biochemical activities.
• Deconvolution to identify the specific open reading frame responsible for an activity.
• Utilizing protein microarrays for high-throughput analysis of immobilized proteins.

Main Results:

• Successfully linked specific biochemical activities to individual genes within the yeast proteome.
• Demonstrated the feasibility of analyzing the entire proteome for enzymatic functions.
• Identified genes responsible for various biochemical activities through a deconvolution process.

Conclusions:

• Genomic protein arrays combined with biochemical assays provide a powerful tool for proteome-wide functional analysis.
• This approach facilitates the systematic identification of gene-function relationships.
• Protein microarrays represent an alternative high-throughput strategy for large-scale protein analysis.