FB2025_01 , released February 20, 2025
Experimental tool: pAPEX
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General Information
Symbol
pAPEX
FlyBase ID
FBto0000405
Name
APEX peroxidase (pea)
Description
Description

pAPEX is an engineered form of the Pisum sativum (pea) ascorbate peroxidase 1 (APX1) gene (UniProtKB:P48534). It can be used as a genetically encoded tag for electron microscopy (EM) or as a proximity labeling tool for proteomic analysis. For EM, cells expressing pAPEX are fixed and then exposed to diaminobenzidine (DAB) and hydrogen peroxide. pAPEX catalyzes the hydrogen peroxide-dependent polymerization and local deposition of DAB, which results in EM contrast after treatment with osmium. This property can be used to study the ultrastructural location of a tagged protein of interest, or to to study the ultrastructure of an organelle or other subcellular structure (PMID:23086203). For proteomic analysis, pAPEX is genetically targeted to a protein complex or organelle of interest and live cells are exposed to hydrogen peroxide and biotin-phenol. pAPEX catalyzes the formation of short-lived biotin-phenoxyl radicals which covalently attach to proximal endogenous proteins, allowing their purification using streptavidin beads (PMID:23371551). pAPEX acts as a monomer and contains the following mutations relative to wild-type pea APX1: K14D, W41F, E112K (K14D and E112K are monomerizing mutations) (PMID:23086203).

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Synonyms and Secondary IDs (5)
Reported As
Symbol Synonym
Name Synonyms
APEX peroxidase
APEX peroxidase (pea)
Secondary FlyBase IDs
    References (5)