title: An efficient method for purification of nonspecific lipid transfer protein-1 from rice seeds using kiwifruit actinidin proteolysis and ion exchange chromatography.
abstract: Plant nonspecific lipid transfer proteins are small basic proteins that transport phospholipids between membranes and are subdivided into two subfamilies, nsLTP(1) (9 kDa) and nsLTP(2) (7 kDa). LTPs have potential application in the defense reactions against pathogens and the drug delivery systems. Many efforts have been made for purification of different nsLTPs from various plants; however, most of them used successive purification procedures. We have developed a relatively simple and efficient method for the purification of rice nsLTP(1), based on the proteolytic activity of kiwifruit actinidin on the rice seed extract and one-step chromatographic procedure on a CM-Sepharose column. The purity of protein was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The isolated LTP(1) migrated as a homogenous polypeptide with molecular mass of 9 kDa that confirms the efficiency of actinidin on the digestion of major contaminations present in the rice seed extract without any harmful effect on the LTP(1). The advantages of using proteolytic activity of actinidin in purifying rice LTP(1) includes the reduced separation time allowing the purification of LTP(1) in one-step chromatographic procedure, low costing, high efficiency, and the relative simplicity of the method.
authors.affiliation: Department of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran.
authors.forename: Sirous
authors.initials: S
authors.lastname: Ghobadi
authors.forename: Fakhroddin
authors.initials: F
authors.lastname: Yousefi
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Khademi
authors.forename: Samira
authors.initials: S
authors.lastname: Padidar
authors.forename: Ali
authors.initials: A
authors.lastname: Mostafaie
mesh_terms: D029042:Actinidia / Q000201:enzymology*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D008970:Molecular Weight; D012275:Oryza / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D059748:Proteolysis; D012639:Seeds / Q000737:chemistry
chemical_list: D002352:Carrier Proteins; D010940:Plant Proteins; C030363:lipid transfer protein; D003546:Cysteine Endopeptidases; C018745:actinidain