Purification and immunocharacterization of a plant cytochrome P450: The cinnamic acid 4-hydroxylase

doi:10.1016/0003-9861(91)90199-S

Archives of Biochemistry and Biophysics

Volume 288, Issue 1, July 1991, Pages 302-309

https://doi.org/10.1016/0003-9861(91)90199-S Get rights and content

Abstract

Cinnamic acid 4-hydroxylase (CA4H) was purified from microsomes of manganese-induced Jerusalem artichoke (Helianthus tuberosus L.) tuber tissues. The three-step purification procedure involved solubilization and phase partitioning in Triton X-114, followed by chromatography on DEAE-Trisacryl and hydroxylapatite columns. Purification was monitored using carbon monoxide and type I substrate binding properties of the enzyme. The protein, purified to electrophoretic homogeneity, showed an M_r of about 57,000 on SDS-PAGE. Polyclonal antibodies raised against this protein selectively reacted with a 57-kDa polypeptide on Western blots of induced Jerusalem artichoke microsomes. The antibody selectively and strongly inhibited CA4H activity from several plant species.

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Purification and immunocharacterization of a plant cytochrome P450: The cinnamic acid 4-hydroxylase

Abstract

J. Biol. Chem

Arch. Biochem. Biophys

J. Biochem

Anal. Biochem

Anal. Biochem

FEBS Lett

J. Colloid Interface Sci

Plant Sci

Phytochemistry

J. Biol. Chem

Anal. Biochem

Anal. Biochem