Main Page/BPHS 4090/Lysozyme Crystallization

Introduction

Suggested Reading:

Introduction to Macromolecular X-Ray Crystallography by Esko Oksanen and Adrian Goldman (Chapter 10 in Comprehensive Natural Products II Chemistry and Biology, Editors-in-Chief: Lew Mander and Hung-Wen (Ben) Liu ISBN: 978-0-08- 045382-8).

Alexander McPherson. Introduction to protein crystallization. Methods, 34, 254-265.

Protein X-Ray Crystallography Methods by Roger S. Rowlett, Department of Chemistry, Colgate University

Websites

• http://www.ruppweb.org/level1/new_tutorials_page.htm
• http://www.sdsc.edu/Xtal/edu.index.html
• http://www.hamptonresearch.com
• http://www.pxuniverse.com/

Experimental

You will be provided with the pure protein (lysozyme) that you will be crystallizing. You will also be provided with the crystallization solution. In real life you perform crystallization trials with your homogeneously pure protein and examine your plates looking for protein crystals.

Crystallization Experiment I

Protein

Lysozyme at 25, 50, 75 and 100 mg/ml dissolved in 0.02 M sodium acetate pH 4.6

Lysozyme Crystallization Solution

• 0.6 M sodium chloride
• 0.1 M sodium acetate pH 4.6
• 25% Glycerol

Procedure

Figure 1: Crystallization Plate

1. Fill reservoirs with 500 μl of lysozyme crystallization solution.
2. Add 1 μl of the crystallization solution to 1 μl of lysozyme on the cover.

Figure 2