ZrO2 Nanofiber as a Versatile Tool for Protein Analysis.

Area of Science:

• Biochemistry
• Proteomics
• Materials Science

Background:

• Phosphorylation is a crucial post-translational modification regulating cellular processes.
• Low abundance of phosphorylated proteins and suppression effects in mass spectrometry necessitate effective enrichment methods.
• Current enrichment techniques often lack efficiency in complex biological samples.

Purpose of the Study:

• To develop and evaluate electrospun zirconium dioxide (ZrO2) nanofibers for enriching phosphorylated peptides and proteins.
• To assess the performance of ZrO2 nanofibers compared to existing commercial enrichment materials.
• To explore the potential of ZrO2 nanofibers for selective protein fractionation based on isoelectric point (pI).

Main Methods:

• Fabrication of ZrO2 nanofibers via electrospinning.
• Application of ZrO2 nanofibers for the enrichment of phosphorylated peptides and proteins from protein digests and cell lysates.
• Comparative analysis of enrichment efficiency against commercial nanoparticles.
• Investigation of pH-dependent selective protein adsorption.

Main Results:

• Electrospun ZrO2 nanofibers exhibited good size homogeneity and porosity.
• The nanofibers demonstrated specific binding to phosphorylated peptides and proteins, enabling effective separation from unmodified species.
• Enrichment performance of ZrO2 nanofibers surpassed that of commercially available nanoparticles.
• Selective adsorption of proteins based on pI was achieved by adjusting solution pH.

Conclusions:

• Electrospun ZrO2 nanofibers provide a versatile and efficient tool for enriching phosphorylated proteins.
• This method enhances the identification and quantification of low-abundance phosphoproteins in complex samples.
• The developed nanofibers offer a promising platform for both phosphoproteomics and charge-based protein fractionation.