Protein methylation identification method

An identification method, methylation technology, which is applied in the identification field of methylated proteome, can solve the problems such as inability to distinguish peptide segments, and achieve the effect of demethylation processing of spectral peaks

Active Publication Date: 2018-08-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But in the traditional labeling method (document 1: S.-E.Ong, G.Mittler, M.Mann, Identifying and quantifying in vivo methylation sites by heavy methyl SILAC, Nat Methods 1 (2004) 119-126), for containing The d

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Protein methylation identification method
  • Protein methylation identification method
  • Protein methylation identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Peak demethylation strategy for large-scale analysis of methylation proteomics: HEK293 cells were cultured in DMEM containing 0.2 mM L‐methionine supplemented with 10% dialyzed fetal bovine serum Culture medium for eight generations (at least eight generations), in the presence of 0.2mM L-methionine-carboxyl- 13 C, Methyl‐D 3 Eight generations (at least eight generations) were cultured in the DMEM medium that added 10% of the fetal bovine serum of the dialyzed treatment; Collect the cells respectively and respectively in the lysate (8M urea, 1% v / v polyethylene glycol octyl phenyl ether , 65mM dithiothreitol, 1mM phenylmethylsulfonyl fluoride, 1% v / v protease inhibitor, 50mM Tris-Cl pH 7.5 buffer solution) in ultrasonic-assisted disruption; Samples and relabeled samples were mixed. Add a final concentration of 20 mM dithiothreitol to 2 mg of protein mixed sample, place in a water bath at 37 °C for 2 h, then add a final concentration of 40 mM iodoacetamide, react in th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a new protein methylation identification method. According to the present invention, the methylation site is labeled by using the novel heavy isotope labeled methionine, and the methylation peptide fragment analysis method integrating the methionine cell culture amino acid stable isotope labeling, the identification and extraction of the fragmentation spectrum of the methylation peptide fragment, and the spectrum peak demethylation treatment is developed, and is used in the large-scale analysis of methylated proteome; the amino acid labeling method can distinguish themethylated peptide fragment from the peptide fragment containing methionine, can identify and extract the fragmentation spectrum derived from the methylated peptide fragment, and can demethylate the fragmentation spectrum so as to obtain the spectrum of the unmethylated peptide fragment, and the peptide fragment sequence is determined by searching in a database so as to determine the modificationsite by binding to the spectrum of the methylated peptide fragment; and the method does not require the preset of the modification type so as to simultaneously analyze all the known methylation types.

Description

technical field [0001] The invention belongs to the technical field of methylated proteomics in the direction of proteomics research, and specifically relates to a new identification method of methylated proteomics. Background technique [0002] Protein methylation is catalyzed by S-adenosylmethionine (SAM)-dependent methyltransferases, and is one of the most common post-translational modifications of proteins. Methylation modification plays an important regulatory role in the regulation of many physiological processes. Studies have shown that protein methylation can regulate the intramolecular or intermolecular interactions of target proteins; affect their affinity with RNA, thereby affecting a variety of cellular processes, including transcription regulation, cellular localization, ribosome assembly, and RNA processing , maturation of heterogeneous ribosomal ribosomal proteins (hnRNPs), protein-protein interactions, accuracy of translation, nuclear transport, protein and ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/68
CPCG01N33/6818
Inventor 叶明亮王科云毛家维
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap