Treatment method of sample for membrane protein analysis identification

Abstract

The present invention relates to the field of membrane proteomics research, specifically to a treatment method of a sample for membrane protein analysis identification. According to the treatment method of the present invention, SL is adopted as a detergent, and is provided for membrane protein analysis identification, wherein SL molecules have a linear hydrophobic hydrocarbon chain tail similar to SDS molecules, and a carboxyl head similar to SDC molecules, and experiment results show that SL provides significant advantages in identifications of membrane proteins, especially in identifications of integral membrane proteins having high hydrophobicity and/or multiple transmembrane regions compared with ALS and SDC.

Description

technical field [0001] The invention relates to the field of membrane proteomics research, in particular to a method for processing samples for analysis and identification of membrane proteins. Background technique [0002] In the study of membrane proteomics, most membrane proteins, especially integral membrane proteins, are highly hydrophobic, which hinders their effective extraction, dissolution and enzymatic hydrolysis, thus affecting their analysis and identification. In order to increase the solubility of membrane proteins, the method of adding solubilizers (such as detergents, organic solvents and denaturants, etc.) to the buffer is usually used to process samples for membrane protein analysis and identification. Compared with other types of solubilizers, detergents are more widely used due to their strong solubilizing ability. When using detergent to extract membrane protein, the concentration of detergent is generally 1-2%. Among them, sodium dodecyl sulfate (SDS)...

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Problems solved by technology

This method can be used for the treatment of samples for membrane protein analysis and identification. However, since the concentration of SDS in the sample is higher than 0.1%, the protease activity will be seriously inhibited. Methods such as dilution, gel-mediated sample purification or ion exchange reduce the concentration of SDS in the sample, but a large dilution will greatly increase the volume of the sample, thereby increasing the amount of enzyme used in enzymatic hydrolysis; and the SDS in the sample is not completely removed, Excessive sample volume also brings difficulties to subsequent analysis and identification; other methods such as gel-mediated sample purification methods or ion exchange are relatively cumbersome to operate and easily cause sample loss

Because the existence of SDS will affect the subsequent protein enzymatic hydrolysis and liquid chromatography-mass spectrometry analysis, which limits its application in sample processing for membrane protein analysis and identification.

Sodium deoxycholate (SDC) and a commercial acid-labile detergent ALS (sodium 3–[(2–methyl–2–undecyl–1,3–dioxolan–4–yl) methoxyl]–1–propanesulfonate , the trade name is RapiGest SF, the literature is generally called ALS or RapiGest SF, there is no Chinese name) is a detergent that has a similar effect to SDS, and they can be used at higher concentrations without affecting the activity of proteases; and because Can be precipitated under acidic conditions and can be easily removed from the sample without interfering with subsequent mass spectrometric analysis; the disadvantage is that they are not as effective at dissolving biofilms and extracting membrane proteins as SDS

In view of the fact that the currently used solubilizers either have poor ability to extract membrane proteins or interfere with subsequent analysis and identification, it is difficult to achieve good results

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