Creating Optimal Conditions for OPA1 Isoforms by Western Blot in Skeletal Muscle Cells and Tissue.

Mitochondria Muscle Tissue Optic atrophy-1 (OPA1) Western Blot isoforms isolation

Journal

bioRxiv : the preprint server for biology
Titre abrégé: bioRxiv
Pays: United States
ID NLM: 101680187

Informations de publication

Date de publication:
19 Aug 2023
Historique:
pubmed: 9 6 2023
medline: 9 6 2023
entrez: 9 6 2023
Statut: epublish

Résumé

OPA1 is a dynamin-related GTPase that modulates various mitochondrial functions and is involved in mitochondrial morphology. There are eight different isoforms of OPA1 in humans and five different isoforms in mice that are expressed as short or long-form isoforms. These isoforms contribute to OPA1's ability to control mitochondrial functions. However, isolating OPA1 all long and short isoforms through western blot has been a difficult task. To address this issue, we outline an optimized western blot protocol to isolate 5 different isoforms of OPA1 on the basis of different antibodies. This protocol can be used to study changes in mitochondrial structure and function.

Identifiants

pubmed: 37292669
doi: 10.1101/2023.05.20.541601
pmc: PMC10245902
pii:
doi:

Types de publication

Preprint

Langues

eng

Auteurs

Dominique C Stephens (DC)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.
Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA.

Margaret Mungai (M)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Amber Crabtree (A)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Heather K Beasley (HK)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Edgar Garza-Lopez (E)

Department of Internal Medicine, University of Iowa, Iowa City, IA, 52242, USA.

Kit Neikirk (K)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Serif Bacevac (S)

Central Microscopy Research Facility, University of Iowa, Iowa City, IA, 52242, USA.

Larry Vang (L)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Zer Vue (Z)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Neng Vue (N)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Andrea G Marshall (AG)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Kyrin Turner (K)

Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA.

Jianqiang Shao (J)

Central Microscopy Research Facility, University of Iowa, Iowa City, IA, 52242, USA.

Sandra Murray (S)

Department of Cell Biology, College of Medicine, University of Pittsburgh, Pittsburgh, PA, 15260, USA.

Jennifer A Gaddy (JA)

Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.
Tennessee Valley Healthcare Systems, U.S. Department of Veterans Affairs, Nashville, TN, 37212, USA.

Celestine Wanjalla (C)

Vanderbilt University Medical Center: Department of Medicine, Division of Infectious Disease, Nashville, TN, USA.

Jamaine Davis (J)

Department of Biochemistry and Cancer Biology. Meharry Medical College, Nashville, TN, 37208, USA.

Steven Damo (S)

Department of Life and Physical Sciences, Fisk University, Nashville, TN, 37232, USA.

Antentor O Hinton (AO)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Classifications MeSH