YOL099C Antibody

YOL099C Overview

YOL099C is categorized as a "dubious" ORF in the Saccharomyces Genome Database (SGD), with no confirmed functional protein product . Key characteristics include:

• Genomic Context: Located on chromosome XV.

• Sequence Features: Lacks homology to conserved protein domains or functional motifs.

• Experimental Evidence: No phenotypes or essentiality reported in knockout studies .

Development and Validation of the YOL099C Antibody

Antibodies against YOL099C have been employed in large-scale proteomic screens to assess the expression of dubious ORFs. Key validation steps include:

Antibody Specificity Testing

• CRISPR/Cas9 Knockout (KO) Validation: Parental and KO yeast strains were compared via immunoblot to confirm antibody specificity. A signal reduction in KO lysates indicates target specificity .

• Glycoprotein Screening: In a yeast proteome study, YOL099C was identified as a candidate glycoprotein using an anti-glycan antibody. The antibody-reactive set showed 4.5-fold enrichment for known glycoproteins, suggesting potential post-translational modification of YOL099C .

Performance Metrics

Evidence of Protein Expression

• A proteome-wide antibody screen detected YOL099C in S. cerevisiae lysates, challenging its "dubious" classification .

• The antibody signal correlated with mRNA abundance data, suggesting low-level expression under standard conditions .

Functional Implications

• Glycosylation Potential: Enrichment in glycoprotein screens implies YOL099C may undergo post-translational modification, though its biological relevance remains unclear .

• Conservation: Absence of orthologs in other fungi supports the hypothesis that YOL099C is non-functional .

Challenges and Limitations

• Specificity Concerns: Cross-reactivity with unrelated proteins was observed in immunoblots, necessitating stringent KO validation .

• Low Abundance: YOL099C protein levels are near detection thresholds, complicating functional studies .

Future Directions

• Structural Studies: Cryo-EM or X-ray crystallography could resolve whether YOL099C forms stable protein structures.

• Conditional Expression: Profiling under stress conditions (e.g., nutrient deprivation) may reveal context-dependent expression.