Method for the characterization of the three-dimensional structure of proteins employing mass spectrometric analysis and computational feedback modeling

• US 20060277017A1
• Filed: 05/05/2006
• Published: 12/07/2006
• Est. Priority Date: 11/04/1993
• Status: Abandoned Application

• Current Assignee: H-Nu Ops, Inc.
• Original Assignee: H-Nu Ops, Inc.
• Inventors: Kruger, Terry L., Sproch, Norman K., Kruger, Karyn
• Application Number:
  US11/418,997
• Publication Number:
  US 20060277017A1
• Time in Patent Office: Days
• US Class Current: 703/12
• CPC Class Codes: G01N 33/6848 Methods of protein analysis involving mass spectrometry
• CPC Class Codes: H01J 49/0431 for liquid samples interfaces to liquid chromatographs G01N30/7233
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

A method for characterizing the three-dimensional surface structure of molecules, particularly proteins and protein complexes, employing mass spectrometric analysis, an electrospray ionization (ES) source, a novel data interpretation process that utilizes comparisons of particular binding constants (K_B) and heats of formation (ΔH_f), and computational feedback modeling.

Claims

1. A method for obtaining spectral data, comprising the steps of:

(a) mixing a small molecule with a large molecule so that the small molecule binds non-covalently to the large molecule to form a large molecule/small molecule complex; and

(b) performing electrospray ionization mass spectrometry to obtain spectral data relating to the large molecule-small molecule complex.

2. The method of claim 1, wherein the large molecule is selected from the group consisting of one or more of polypeptides, proteins, DNA, RNA, and oligosaccharides.

3. The method of claim 1, wherein the large molecule comprises a protein molecule.

4. The method of claim 1, wherein the small molecule is selected from the group consisting of crown ethers, cryptands, and macrocyclic polyethers.

5. The method of claim 1, wherein the small molecule comprises a crown ether.

6. A method for obtaining spectral data, comprising the steps of:

(a) mixing a crown ether with a large molecule so that the crown ether binds non-covalently to the large molecule to form a large molecule/crown ether complex; and

(b) performing electrospray ionization mass spectrometry to obtain spectral data relating to the large molecule/crown either complex.

7. The method of claim 6, wherein the large molecule is selected from the group consisting of one or more of polypeptides, proteins, DNA, RNA, and oligosaccharides.

8. The method of claim 6, wherein the large molecule comprises a protein molecule.