Heme o Isolation and Analysis Using Heme o Synthase Heterologously Expressed in Escherichia coli.
Methods Mol Biol
; 2839: 131-149, 2024.
Article
em En
| MEDLINE
| ID: mdl-39008252
ABSTRACT
Heme o is an Fe-porphyrin involved in the majority of aerobic respiration pathways found in all three domains of life. In eukaryotes and most aerobic prokaryotes, heme o functions solely as the precursor for the synthesis of heme a, a necessary cofactor for most heme-copper terminal oxidases. In some prokaryotes, such as Escherichia coli (E. coli), heme o can serve as a cofactor for heme-copper oxidases instead of heme a. Given its role as a key substrate or cofactor, purified heme o promises to be a valuable resource for the study of heme-copper oxidase assembly and activity. However, commercially available heme o is sold in limited quantities at a relatively high cost (compared to the prototypical heme b), making the use of heme o purchased from suppliers unfeasible for such studies. In this chapter, we present step-by-step methods both for heme o isolation from E. coli overexpressing heme o synthase and for HPLC analysis of cellular hemes (i.e., heme o and heme b).
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01-internacional
Base de dados:
MEDLINE
Assunto principal:
Escherichia coli
/
Heme
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article