Hydrophilic thiol probe

Patent metadata

Field	Value
Publication number	US-9448240-B2
Application number	US-201214241728-A
Country	US
Kind code	B2
Filing date	Aug 17, 2012
Priority date	Sep 9, 2011
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

Abstract

A thiol probe for a protein, which promotes ionization in proteomic analysis using mass spectrometry and can be used even for a protein that has a high degree of hydrophobicity and quickly turns over, and is represented by the following formula (I):

In formula (I), R 1 represents a linker group, and R 2 represents a substituted ammonium group or a substituted amino group. A mass spectrometry method for a protein, includes the steps of obtaining a modified protein by reacting the thiol probe with a protein to be subjected to mass spectrometry and subjecting the modified protein to mass spectrometry.

First claim

The invention claimed is:

1. A thiol probe for a protein, which is represented by the following formula (I):

wherein R 1 represents a linker group which is either a hydrocarbon group having 1 to 3 carbon atoms or an alkylene oxide-containing group having 2 to 6 carbon atoms, and wherein an alkylene oxide in the alkylene oxide-containing group is ethylene oxide or propylene oxide, and
wherein R 2 represents a substituted amino group represented by —NHR 3 , wherein R 3 represents an amidino group which may be substituted, or a triazino group which may be substituted.

2. The thiol probe for a protein according to claim 1 , wherein a substituent group in the triazino group which may be substituted is selected from the group consisting of an amino group and an alkoxy group having 1 or 2 carbon atoms.

3. The thiol probe for a protein according to claim 2 , which is represented by the following formula (vi):

4. The thiol probe for a protein according to claim 2 , which is represented by the following formula (vii):

5. The thiol probe for a protein according to claim 1 , wherein a substituent group in the amidino group which may be substituted is an alkyl group having 1 or 2 carbon atoms.

6. The thiol probe for a protein according to claim 1 , which is represented by the following formula (iv):

7. The thiol probe for a protein according to claim 1 , which is represented by the following formula (v):

8. A thiol probe for a protein, which is represented by the following formula (I):

wherein R 1 represents a linker group which is either a hydrocarbon group having 1 to 3 carbon atoms or an alkylene oxide-containing group having 2 to 6 carbon atoms, and wherein an alkylene oxide in the alkylene oxide-containing group is ethylene oxide or propylene oxide, and
wherein R 2 represents a substituted ammonium group, wherein the substituted ammonium group is a tertiary or quaternary ammonium group substituted by an alkyl group having 1 or 2 carbon atoms.

9. The thiol probe for a protein according to claim 8 , which is represented by the following, formula (i):

10. The thiol probe for a protein according to claim 8 , which is represented by the following formula (ii):

11. The thiol probe for a protein according to claim 8 , which is represented by the following, formula (iii):

12. A mass spectrometry method for a protein, comprising the steps of:
obtaining a modified protein by reacting a thiol probe for a protein with a protein; and
subjecting the modified protein to mass spectrometry,
wherein the thiol probe for a protein is represented by the following formula (I):

wherein R 1 represents a linker group, and R 2 represents a substituted ammonium group or a substituted amino group,
wherein the linker group is an alkylene oxide-containing group having 2 to 6 carbon atoms, and
wherein an alkylene oxide in the alkylene oxide-containing group is ethylene oxide or propylene oxide.

13. A mass spectrometry method for a protein, comprising the steps of:
obtaining a modified protein by reacting a thiol probe for a protein with a protein; and
subjecting the modified protein to mass spectrometry,
wherein the thiol probe for a protein is represented by the following formula (I):

wherein R 1 represents a linker group, and R 2 represents a substituted amino group, and
wherein the substituted amino group is a group represented by —NHR 3 , wherein R 3 represents a hydrocarbon group or a nitrogen-containing group.

14. The mass spectrometry method for a protein according to claim 13 , wherein R 3 is an amidino group which may be substituted, or a triazino group which may be substituted.

15. The mass spectrometry method for a protein according to claim 14 , wherein a substituent group in the triazino group which may be substituted is selected from the group consisting of an amino group and an alkoxy group having 1 or 2 carbon atoms.

16. The mass spectrometry method for a protein according to claim 15 , wherein the thiol probe is represented by the following formula (vi):

17. The mass spectrometry method for a protein according to claim 15 , wherein the thiol probe is represented by the following formula (vii):

18. The mass spectrometry method for a protein according to claim 14 , wherein a substituent group in the amidino group which may be substituted is an alkyl group having 1 or 2 carbon atoms.

19. The mass spectrometry method for a protein according to claim 14 , wherein the thiol probe is represented by the following formula (iv):

20. The mass spectrometry method for a protein according to claim 14 , wherein the thiol probe is represented by the following formula (v):

21. A mass spectrometry method for a protein, comprising the steps of:
obtaining a modified protein by reacting a thiol probe for a protein with a protein; and
subjecting the modified protein to mass spectrometry,
wherein the thiol probe for a protein is represented by the following formula (I):

wherein R 1 represents a linker group, and R 2 represents a substituted ammonium group, and
wherein the substituted ammonium group is a tertiary or quaternary ammonium group substituted by an alkyl group having 1 or 2 carbon atoms.

22. The mass spectrometry method for a protein according to claim 21 , wherein the thiol probe is represented by the following formula (i):

23. The mass spectrometry method for a protein according to claim 21 , wherein the thiol probe is represented by the following formula (ii):

24. The mass spectrometry method for a protein according to

Assignees

• Shimada Takashi
• Sato Taka-Aki
• Tanaka Koichi
• Shimadzu Corp

Inventors

• Shimada Takashi
• Sato Taka-Aki
• Tanaka Koichi

Classifications

• Y10T436/19

  Halogen containing · CPC title

• Y10T436/145555

  Hetero-N · CPC title

• Y10T436/147777

  Plural nitrogen in the same ring [e.g., barbituates, creatinine, etc.] · CPC title

• Y10T436/24

  Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry · CPC title

• G01N33/6848 Primary

  Methods of protein analysis involving mass spectrometry · CPC title