Optimization of protein extraction from Hypericum perforatum tissues and immunoblotting detection of Hyp-1 at different stages of leaf development.

Abstract:

:Sample preparation is crucial for obtaining high-quality proteins for the purpose of electrophoretic separation and further analysis from tissues that contain high levels of interfering compounds. Hypericum perforatum is a medicinal plant that contains high amounts of phenolic compounds, of which hypericins, hyperforins, and flavonoids contribute to the antidepressant activities of the plant. This study focuses on obtaining optimized amounts of high-quality proteins from H. perforatum, which are suitable for electrophoretic analyses. From the tested protein extraction solutions, sodium borate buffers at pH 9 and 10 gave the best protein yields from mature H. perforatum leaves. With these buffers, relatively high protein yields could also be obtained from roots, stems, and flower buds. The protein extracts of all organs were well resolved in SDS-PAGE after an efficient removal of non-protein contaminants with PVPP, phenol extraction, and methanolic ammonium acetate precipitation. The method was suitable for high-quality protein extraction also from other tested species of genus Hypericum. The applicability of the protocol for immunoblotting was demonstrated by detecting Hyp-1 in H. perforatum leaves at different stages of development. Hyp-1, which has been suggested to attend to the biosynthesis of hypericin, accumulated in high amounts in H. perforatum leaves at mature stage.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Karppinen K,Taulavuori E,Hohtola A

doi

10.1007/s12033-010-9299-9

subject

Has Abstract

pub_date

2010-11-01 00:00:00

pages

219-26

issue

3

eissn

1073-6085

issn

1559-0305

journal_volume

46

pub_type

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