Analyzing the integrity of oxidative phosphorylation complexes in Drosophila flight muscles

STAR Protoc. 2021 Dec 13;2(4):101021. doi: 10.1016/j.xpro.2021.101021. eCollection 2021 Dec 17.

Abstract

Drosophila flight muscles are highly enriched with mitochondria and have emerged as a powerful genetic system for studying how oxidative phosphorylation (OXPHOS) complexes are assembled. Here, we describe a series of protocols for analyzing the integrity of OXPHOS complexes in Drosophila via blue native polyacrylamide gel electrophoresis (BN PAGE). We have also included protocols for the additional steps that are typically performed after OXPHOS complexes are separated by BN PAGE, such as Coomassie staining, silver staining, and in-gel OXPHOS activities. For complete details on the use and execution of this protocol, please refer to Murari et al. (2020).

Keywords: Cell Biology; Genetics; Metabolism; Model Organisms; Protein Biochemistry.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Drosophila Proteins* / analysis
  • Drosophila Proteins* / chemistry
  • Drosophila Proteins* / metabolism
  • Drosophila* / chemistry
  • Drosophila* / physiology
  • Female
  • Flight, Animal / physiology*
  • Male
  • Muscle Proteins* / analysis
  • Muscle Proteins* / chemistry
  • Muscle Proteins* / metabolism
  • Muscles* / chemistry
  • Muscles* / metabolism
  • Native Polyacrylamide Gel Electrophoresis
  • Oxidative Phosphorylation

Substances

  • Drosophila Proteins
  • Muscle Proteins