Phosphoproteomic Analysis of Yeast

Phosphorylation is one of the most common post-translational modifications of proteins and is the most important form of biological regulatory modification. Protein phosphorylation is formed by the transfer of phosphate groups on ATP or GTP molecules to specific amino acid side chains of substrate proteins under the action of protein kinases. Protein phosphorylation can participate in regulating the growth, development, adversity stress, disease occurrence and other life processes of organisms by stimulating and regulating many signaling pathways. Therefore, phosphorylation has always been the focus and hotspot of biological research. Mass spectrometry-based quantitative proteomic studies of phosphorylation include the qualitative and quantitative analysis of phosphoproteom, providing detailed information on identified phosphorylation sites. 

Due to the low content and wide dynamic range of phosphorylated proteins in biological samples, it is necessary to enrich for the modifications to increase the abundance of phosphorylated proteins before quantitative analysis of phosphorylated peptide samples. In mass spectrometry-based quantitative proteomic studies of phosphorylation, phosphorylated proteins can be enriched using solid-phase metal affinity chromatography, phosphorylated antibodies or TiO2. And in the process of enriching the peptides, the monophosphorylated peptides and the polyphosphorylated peptides are separated step by step, and finally the mass spectrometry analysis is carried out. 

Service introduction

Creative BioMart uses mass spectrometry platform (Q Exactive HF, Orbitrap Fusion, Orbitrap Fusion Lumos) combined with nano LC - MS / MS to launch a service package for yeast phosphoproteomic analysis. You only need to tell us your experimental purpose and send us your samples, and we will take care of all the follow-up matters, including yeast protein extraction, protease digestion, phosphorylated peptide enrichment, peptide separation, MS analysis and bioinformatics analysis. If you have any needs in this regard, please feel free to contact us.

Provided by clients

• Yeast cell number ≥ 5 × 10⁷ 
• Yeast ≥ 200 mg (dry weight)
• Yeast protein extract ≥ 1 mg, concentration > 2 mg / ml.

Samples should not contain components that would interfere with separation, such as nucleic acids, lipids and polysaccharides.

• Sample information sheet (the source, content, status and other basic information of the sample).

Experiment procedure

Delivery content

Creative BioMart will provide you with detailed technical reports including:

• Experimental procedure
• Relevant mass spectrometry parameters
• Details of the identified phosphorylation sites
• Mass spectrometry
• Raw data

Project cycle

• The operating cycle of the standard process is 45 working days.

Service process

Creative BioMart specializes in quantitative proteomics services for yeast post-translational modifications. Our researchers have many years of experience in this field. They can quickly understand the project situation through a simple description, and quickly formulate a feasible research plan according to customer needs. If you have this need, please feel free to contact us.