Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methods for analysing protein samples based on the identification of c-terminal peptides

A carboxy-terminal peptide and protein technology, applied in the field of data analysis of technology and mass spectrometry data, can solve the problem that peptides cannot be attributed and achieve the effect of reducing complexity

Inactive Publication Date: 2009-08-26
KONINKLIJKE PHILIPS ELECTRONICS NV
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, considering the fact that the analysis of the complete sample still yields numerous different peptides with the same mass and the same physicochemical properties, such peptides cannot be attributed to a single parent protein

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
  • Methods for analysing protein samples based on the identification of c-terminal peptides
  • Methods for analysing protein samples based on the identification of c-terminal peptides
  • Methods for analysing protein samples based on the identification of c-terminal peptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] Example 1: Separation of carboxy-terminal peptides

[0140] The flow chart for the isolation of carboxy-terminal peptides is at figure 1 in brief.

[0141] Isolate protein extracts from tissues using standard methods. The side chain of cysteine ​​is alkylated and the amine at the amino terminus and the side chain of lysine are acetylated. In the next step, 1-ethyl-3-[3-dimethylamino-propyl]carbodiimide hydrochloride was added according to the method as shown in Grabarek and Gergely (1990) Anal Biochem. (EDC) or 1-ethyl-3(3-dimethyl-aminopropyl)-carbodiimide (EDAC) activates the free carboxyl group of the carboxy-terminal amino acid (and the reactivity on glutamate and aspartate carboxyl).

[0142] EDC and protein ( figure 2 The carboxyl group on molecule 1) in reacts to form an amine-reactive O-acylisourea intermediate. This intermediate can be combined with NH-R( figure 2 The amine reaction on molecule 2) in , produces a conjugate of these two molecules linked...

Embodiment 2

[0147] Example 2: Identification of peptides with similar masses

[0148] This example shows the need to supplement peptide quality data with additional parameters. From Prosite(ScanProsite on www.expasy.org / prosite), the sequence SFPNIGSL [SEQ ID NO: 1] of an 8-amino acid carboxy-terminal tryptic peptide of a human protein with potential clinical relevance, Exostosin 2, was selected.

[0149] The calculated average mass (833.94) of SEQ ID NO: 1 was used to identify Profound (prowl.rockefeller.edu) peptides with calculated masses in the range of theoretical 1 Da. This calculation was performed by performing an in silico trypsinization of human protein (see Table 1 ) which does not allow partial cleavage and does not select a large number of peptides which are carboxy-terminal.

[0150] Table 1. Masses and physicochemical parameters of carboxy-terminal peptides with relevant masses

[0151] 1

2

3

4

5

6

7

quality

...

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 present invention relates to a method for identifying proteins in one or more samples based on the isolation and analysis of their C-terminal peptides. The isolated peptides are purified and analysed by Mass spectroscopy. Identification of the parent protein is based on the mass of the C-terminal peptide in combination with additional physicochemical parameters. The present invention further relates to an annotated database of C-terminal peptides of in silico cleaved proteins comprising the masses of C-terminal peptides and one or more physicochemical properties thereof.

Description

field of invention [0001] The present invention relates to a method for the simultaneous analysis of protein samples using mass spectrometry, which method allows the selective isolation of peptides from a mixture of cleaved proteins. The invention further relates to techniques for purification of peptides and data analysis of mass spectral data. Background of the invention [0002] Over the past decades, different methods of identifying proteins using mass spectrometry (MS) have been developed. In so-called fingerprinting, proteins are separated and cleaved into peptide fragments. By comparing the mass of the peptide produced with an in silico database of cleaved proteins, the parent protein can be identified without further sequencing. [0003] However, the purpose of the present invention is to use the shotgun method to study different proteins present in a sample in a single test. Peptide sample complexity is often reduced by selectively isolating peptides containing i...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848
Inventor R·霍夫曼
Owner KONINKLIJKE PHILIPS ELECTRONICS NV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com