Structure of beta-purothionin in membranes: a two-dimensional infrared correlation spectroscopy study.
Two-dimensional infrared correlation spectroscopy has been used to investigate the structure of beta-purothionin, a small basic protein found in the endosperm of wheat seeds, in the absence and presence of dimyristoylphosphatidylglycerol (DMPG) membranes. To generate the two-dimensional synchronous and asynchronous maps, hydrogen-deuterium exchange of the protein amide protons has been used as an external perturbation. This method has allowed us to separate the different secondary structure elements and side chain contributions in the regions of amide I, II, and II' bands to determine that the relative order of deuteration of the beta-purothionin protons is as follows: turns, asparagines, and lysines > unordered structure and tyrosine > beta-sheet > alpha-helices and arginines. The results also indicate that the protein undergoes significant changes both in secondary structure and in deuteration in the presence of DMPG bilayers. The helical content of beta-purothionin is higher in the presence of the lipid, and the relative order of deuteration is as follows: lysines and arginines > asparagines and beta-sheet > unordered structure and alpha-helices. The inversion in the deuteration order of the arginine residues is assigned to a change of the degree of association of the protein in the membrane. In addition, the results reveal that the part of the protein containing the tyrosine residue interacts with the lipid membrane. Our results combined with those previously published suggest that the toxicity of beta-purothionin is more associated with the formation of functional channels in cell membranes rather than with a lytic phenomenon.
Richard
Julie-Andrée
JA
Département de chimie, Centre de Recherche en Sciences et Ingénierie des Macromolécules, Université Laval, Québec, Canada G1K 7P4.
Kelly
Isabelle
I
Marion
Didier
D
Auger
Michèle
M
Pézolet
Michel
M
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides; D003903:Deuterium; D007700:Kinetics; D008958:Models, Molecular; D010715:Phosphatidylglycerols / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012639:Seeds / Q000737:chemistry; D013055:Spectrophotometry, Infrared; D014118:Toxins, Biological / Q000737:chemistry; D014908:Triticum / Q000737:chemistry*
D023181:Antimicrobial Cationic Peptides; D010715:Phosphatidylglycerols; D010940:Plant Proteins; D014118:Toxins, Biological; C016085:purothionin; D003903:Deuterium; C002773:dimyristoylphosphatidylglycerol