Purification of Lipid Transfer Protein 2 (LTP2) from Iranian rice paddy

Journal of Cell and Molecular Research

Article 1, Volume 1, Issue 2, January 0, Pages 72-76   

DOI: 10.22067/jcmr.v1i2.3222

Authors

Mehran Miroliaei^* ; Samira Padidar; Ali Moیtafaie; Sirous Ghobad

Razi University

Abstract

Plant nonspecific lipid transfer proteins (nsLTPs) are divided into nsLTP1 and nsLTP2. The existence of an
internal hydrophobic cavity, is a typical characteristic of nsLTPs that serves as the binding site for lipid
substrates. In this communication a simple, rapid and low-cost alternative method was developed for
purification of nsLTP2 from rice paddy. After extracting, final supernatant was loaded on CM-Sepharose
column, which had previously equilibrated with 0.05 M Tris-HCl buffer, pH 8. Bounded proteins were
separated using a linear gradient of 0-0.5 M NaCl. Solution of separated proteins was dialyzed and applied on a
Phenyl-Sepharose column which previously equilibrated with Tris-HCl 0.05 M, ammonium sulfate 1.5 M,
EDTA 0.005 M and NaHSO3 0.3%, pH 8.4. Tris-Tricin SDS-PAGE of separated proteins, obtained from ionexchange
column, showed multiple bands in the range of 2-20 kDa. Further purification using hydrophobic
column resulted in single band of nsLTP2 at about 7 kDa, reflecting a purified sample in the gel.
Keywords: purification, plant lipid transfer proteins, cation-exchange chromatography, hydrophobic
chromatography

Keywords

purification; plant lipid transfer proteins; cation-exchange chromatography; hydrophobic chromatography