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Protein Separation Using Immobilized Phospholipid
Chromatography
Tzong-Hsien Lee and Marie-Isabel Aguilar
Summary
The chromatographic support containing monolayers of phospholipids offers novel
modes in analyzing and separating proteins. The polar choline head groups on immobilized
phosphatidylcholine were used for the affinity purification of phospholipase A (PLA). The
purification process involves removing the contaminating proteins with detergent additives
to the elution buffer such as short-chain alkylsulfonates. The lipid-bound PLA was eluted
with acetonitrile or octyllysophosphatidylcholine. The purity of PLA was approximately
70% based on densitometric scans of gel electrophoresis. These results suggest that the
lipid-immobilized chromatography may be applied to develop purification methods for
PLA, enzymes, and membrane proteins obtained from diverse cells.
Key Words: Immobilized lipid chromatography; membrane proteins; detergent;
organic solvent.
1. Introduction
Analysis of genomic sequence data estimated that 30% of the proteins derived
from Homo sapiens, Escherichia coli, and Saccharomyces cerevisae will be
integral membrane proteins (1–3). However, while the number of predicted
gene sequences for integral membrane proteins has increased over the last few
years, there is considerably less information about their structure and the nature
of their function within the membrane.
From: Methods in Molecular Biology, vol. 421: Affinity Chromatography: Methods and Protocols, Second Edition
Edited by: M. Zachariou © Humana Press, Totowa, NJ
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