Purification and characterization of the 7S vicilin from Korean pine (Pinus koraiensis).
Pine nuts are economically important as a source of human food. They are also of medical importance because numerous pine nut allergy cases have been recently reported. However, little is known about the proteins in pine nuts. The purpose of this study was to purify and characterize pine nut storage proteins. Reported here is the first detailed purification protocol of the 7S vicilin-type globulin from Korean pine (Pinus koraiensis) by gel filtration, anion exchange, and hydrophobic interaction chromatography. Reducing SDS-PAGE analysis indicated that purified vicilin consists of four major bands, reminiscent of post-translational protease cleavage of storage proteins during protein body packing in other species. The N-terminal ends of vicilin peptides were sequenced by Edman degradation. Circular dichroism (CD) and differential scanning calorimetry (DSC) analyses revealed that pine nut vicilin is stable up to 80 degrees C and its folding-unfolding equilibrium monitored by intrinsic fluorescence can be interpreted in terms of a two-state model.
Jin
Tengchuan
T
Department of Biological, Chemical, and Physical Sciences, Illinois Institute of Technology, Chicago, Illinois 60616, USA.
Albillos
Silvia M
SM
Chen
Yu-Wei
YW
Kothary
Mahendra H
MH
Fu
Tong-Jen
TJ
Zhang
Yu-Zhu
YZ
D000595:Amino Acid Sequence; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D028223:Pinus / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011489:Protein Denaturation; D017510:Protein Folding; D055314:Seed Storage Proteins; D012639:Seeds / Q000737:chemistry; D013816:Thermodynamics
D010446:Peptide Fragments; D010940:Plant Proteins; D055314:Seed Storage Proteins; C015575:vicilin protein, plant