Method of magnetically aligning and crystallizing membrane proteins

Patent metadata

Field	Value
Publication number	US-10688442-B2
Application number	US-201615741498-A
Country	US
Kind code	B2
Filing date	Jul 5, 2016
Priority date	Jul 2, 2015
Publication date	Jun 23, 2020
Grant date	Jun 23, 2020

Abstract

The disclosure describes a method of forming highly ordered membrane protein crystals. The forming process is done in the presence of a magnetic field to exploit the diamagnetic anisotropy of the membrane protein. Further described is a method of magnetic alignment and crystallization of membrane proteins in two-dimensional (2D) sheets for protein structural characterization and applications in functional devices. Block co-copolymers are used in alternative embodiments to assist with the crystallization process.

First claim

What is claimed is:

1. A method of forming a two-dimensional membrane protein crystal comprising:
providing a solution containing a membrane protein and a block co-polymer;
exposing the solution to a magnetic field to facilitate alignment of the membrane protein in the solution; and
forming a membrane protein crystal from the solution, wherein the membrane protein crystal is formed in the presence of the magnetic field.

2. The method of claim 1 , wherein forming a membrane protein crystal from the solution comprises:
concentrating the solution through dialysis.

3. The method of claim 1 , wherein a strength of the magnetic field is based on a diamagnetic anisotropy of the membrane protein.

4. The method of claim 1 , wherein a strength of the magnetic field is based on a diamagnetic anisotropy of the block co-polymer.

5. The method of claim 3 or 4 , wherein the diamagnetic anisotropy is estimated from a molecular simulation model.

6. The method of claim 3 or 4 , wherein the diamagnetic anisotropy is the sum of the diamagnetic anisotropy parallel to the magnetic field and the diamagnetic anisotropy perpendicular to the magnetic field.

7. The method of claim 1 , wherein a difference between a magnetic energy parallel to the magnetic field and a magnetic energy perpendicular to the magnetic field is greater than a thermal energy of the solution.

8. The method of claim 1 , wherein a mass fraction of the block co-polymer in the solution is less than 10%.

9. The method of claim 8 , wherein a diamagnetic anisotropy of the block co-polymer is larger than a diamagnetic anisotropy of the membrane protein.

10. A product created from the process of claim 1 .

11. A functional device incorporating a crystallized membrane protein comprising:
a support layer having a plurality of pores;
a membrane protein crystal formed in the presence of a magnetic field, wherein a diameter of the membrane protein crystal is larger than a diameter of the plurality of holes; and
a bonding agent between the support layer and the membrane protein crystal.

12. The functional device of claim 11 , wherein the bonding agent is a functionalized group on at least one of the support layer and the membrane protein crystal.

Assignees

• Univ Carnegie Mellon
• The Pennsylvania State Univ
• Penn State Res Found

Inventors

• Mauter Meagan
• Kumar Manish

Classifications

• B01D71/261

  Polyethylene · CPC title

• B01D69/10

  Supported membranes; Membrane supports · CPC title

• B01D69/06

  Flat membranes · CPC title

• B01D67/00046 Primary

  by deposition by filtration through a support or base layer · CPC title

• B01D2323/35

  Use of magnetic or electrical fields · CPC title