CYP71A13 Antibody

Introduction to CYP71A13 Antibody

CYP71A13 is a cytochrome P450 monooxygenase enzyme critical for camalexin biosynthesis in Arabidopsis thaliana, a defense compound against necrotrophic pathogens like Alternaria brassicicola and Botrytis cinerea. The CYP71A13 antibody refers to immunological tools (e.g., polyclonal or monoclonal antibodies) designed to detect and study this enzyme’s expression, localization, and interactions in plant research. While direct commercial availability of CYP71A13-specific antibodies is not explicitly documented in the provided sources, its role in biochemical pathways and regulatory networks is well-established through genetic, biochemical, and co-immunoprecipitation (co-IP) studies.

2.1. Enzymatic Role in Camalexin Biosynthesis

CYP71A13 catalyzes the conversion of indole-3-acetaldoxime (IAOx) to indole-3-acetonitrile (IAN), a pivotal step in camalexin production . This reaction is critical for channeling intermediates into the camalexin pathway and avoiding toxic side products.

• CYP71A13 interacts with CYP71B15 (PAD3) and CYP79B2 (Trp conversion enzyme) in a metabolon complex, ensuring efficient substrate channeling .

• CYP71A13 also interacts with Arabidopsis P450 Reductase1 (ATR1), a cytochrome P450 reductase essential for electron transfer .

2.2. Genetic and Pathogen-Response Studies

• Camalexin deficiency: cyp71a13 mutants show reduced camalexin production and increased susceptibility to A. brassicicola .

• Co-regulation with PAD3: CYP71A13 and PAD3 are transcriptionally co-regulated by WRKY33, a transcription factor activated during pathogen infection .

3.1. Co-Immunoprecipitation (Co-IP) and Localization

• Protein interactions: CYP71A13 co-purifies with CYP71B15, CYP71A12, and GSTU4 in microsomal fractions, as shown through co-IP and FRET-FLIM .

• Subcellular localization: CYP71A13-GFP fusion proteins localize to the endoplasmic reticulum (ER), co-localizing with ER markers (e.g., RFP-HDEL) and other P450 enzymes .

3.2. Enzymatic Parameters in Heterologous Systems

• Yeast expression: CYP71A13 converts IAOx to IAN with NADPH-dependent activity. In the presence of glutathione, side products like GS-IAN are formed .

• Kinetic parameters: Turnover rates and substrate affinity (K_m) for CYP71A13 and its homologs (e.g., CYP71A12) are measured via HPLC and Michaelis-Menten kinetics .

Antibody Applications in CYP71A13 Research

While the provided sources do not explicitly describe commercial CYP71A13 antibodies, antibodies for related proteins (e.g., CYP71B15, WRKY33) are used in immunoprecipitation, Western blotting, and chromatin immunoprecipitation (ChIP) assays . For CYP71A13-specific antibodies, researchers likely employ:

1. Polyclonal antibodies: Raised against recombinant CYP71A13 protein for Western blot detection.

2. Tagged fusion proteins: GFP/YFP-tagged CYP71A13 constructs enable pull-down assays and fluorescence microscopy .

5.1. Transcriptional Regulation

• WRKY33: Binds directly to the PAD3 promoter and indirectly regulates CYP71A13 during pathogen-induced camalexin biosynthesis .

• MPK4: A MAP kinase upstream of WRKY33, linking stress signaling to camalexin production .

5.2. Crosstalk with Glucosinolate Pathways

• CYP83A1: Competes with CYP71A13 for IAOx, influencing glucosinolate and camalexin balances . Mutants lacking CYP83A1 accumulate toxic intermediates (e.g., 5-MPTO) but exhibit enhanced resistance to powdery mildew .