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Literature DB >> 35188666

In Vivo Epitope Tagging of Plant Mitochondria.

Abstract

Mitochondria play a key role in cellular metabolism. Analyses of the genome, the proteome, metabolic, physiological, and biochemical functions of mitochondria frequently require the isolation of intact and functional mitochondria from various plant tissues with sufficient yield. For this purpose, we generated a transgenic Arabidopsis thaliana (Arabidopsis) line which presents a triple hemagglutinin tag on the surface of the outer mitochondrial membrane. The affinity tag enables immunocapture of the organelles in a single step. This chapter gives detailed instructions on how to generate transgenic Arabidopsis lines harboring a ubiquitously expressed 3xHA-sGFP-TOM5 mitochondrial fusion protein that is targeted to the outer mitochondrial membrane and enables purification of the organelles in a single step.

Entities: Chemical

Keywords: Arabidopsis; HA-tag; Immunocapture; Magnetic beads; Mitochondria; Organelle isolation; TOM5

Mesh：

Substances：

Year: 2022 PMID： 35188666 DOI： 10.1007/978-1-0716-1791-5_14

Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745

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References

16 in total

1. Analysis of the Arabidopsis mitochondrial proteome.

Authors: A H Millar; L J Sweetlove; P Giegé; C J Leaver
Journal: Plant Physiol Date: 2001-12 Impact factor: 8.340

2. Purification and characterization of the preprotein translocase of the outer mitochondrial membrane from Arabidopsis. Identification of multiple forms of TOM20.

Authors: W Werhahn; A Niemeyer; L Jänsch; V Kruft; U K Schmitz; H Braun
Journal: Plant Physiol Date: 2001-02 Impact factor: 8.340

3. Targeting and assembly of mitochondrial tail-anchored protein Tom5 to the TOM complex depend on a signal distinct from that of tail-anchored proteins dispersed in the membrane.

Authors: Chika Horie; Hiroyuki Suzuki; Masao Sakaguchi; Katsuyoshi Mihara
Journal: J Biol Chem Date: 2003-08-01 Impact factor: 5.157

Review 4. The protein import machinery of mitochondria.

Authors: Nils Wiedemann; Ann E Frazier; Nikolaus Pfanner
Journal: J Biol Chem Date: 2004-02-18 Impact factor: 5.157

5. Tom5 functionally links mitochondrial preprotein receptors to the general import pore.

Authors: K Dietmeier; A Hönlinger; U Bömer; P J Dekker; C Eckerskorn; F Lottspeich; M Kübrich; N Pfanner
Journal: Nature Date: 1997-07-10 Impact factor: 49.962

6. MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo.

Authors: Erol C Bayraktar; Lou Baudrier; Ceren Özerdem; Caroline A Lewis; Sze Ham Chan; Tenzin Kunchok; Monther Abu-Remaileh; Andrew L Cangelosi; David M Sabatini; Kıvanç Birsoy; Walter W Chen
Journal: Proc Natl Acad Sci U S A Date: 2018-12-12 Impact factor: 11.205

In Vivo Epitope Tagging of Plant Mitochondria.

1. Analysis of the Arabidopsis mitochondrial proteome.

2. Purification and characterization of the preprotein translocase of the outer mitochondrial membrane from Arabidopsis. Identification of multiple forms of TOM20.

3. Targeting and assembly of mitochondrial tail-anchored protein Tom5 to the TOM complex depend on a signal distinct from that of tail-anchored proteins dispersed in the membrane.

Review 4. The protein import machinery of mitochondria.

5. Tom5 functionally links mitochondrial preprotein receptors to the general import pore.

6. MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo.

7. Affinity purification of cell-specific mitochondria from whole animals resolves patterns of genetic mosaicism.

8. Rapid Single-Step Affinity Purification of HA-Tagged Plant Mitochondria.

9. Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism.

10. Isolation of intact, functional mitochondria from the model plant Arabidopsis thaliana.