Abstract
Biogenesis of the mitochondrial respiratory chain enzymes involves the coordinated action of the mitochondrial and nuclear genomes. As a matter of fact, the structural subunits forming these multimeric enzymes are encoded in both genomes. In addition, the assistance of nuclear encoded factors, termed assembly factors, is necessary to allow for the expression of the mitochondrial DNA-encoded subunits and to facilitate their maturation, membrane insertion, and further assembly into the corresponding enzymatic complex. These processes involve transient interactions among the newly synthesized mitochondrial products and specific assembly factors. The identification and characterization of these interactions can be achieved by the method described here, consisting of pulling down tagged versions of the assembly factors immediately after radiolabeling the mitochondrial translation products in isolated mitochondria, and analyzing the radiolabeled pulled-down material.
| Original language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Pages | 125-139 |
| Number of pages | 15 |
| Volume | 457 |
| DOIs | |
| State | Published - Dec 1 2008 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 457 |
| ISSN (Print) | 10643745 |
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Keywords
- GST pull-down
- In organello protein synthesis
- Mitochondria; mitochondrial biogenesis
- Mitochondrial translation
- Saccharomyces cerevisiae
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Exploring protein-protein interactions involving newly synthesized mitochondrial dna-encoded proteins. / Horn, Darryl; Fontanesi, Flavia; Barrientos, Antoni.
Methods in Molecular Biology. Vol. 457 2008. p. 125-139 (Methods in Molecular Biology; Vol. 457).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Exploring protein-protein interactions involving newly synthesized mitochondrial dna-encoded proteins
AU - Horn, Darryl
AU - Fontanesi, Flavia
AU - Barrientos, Antoni
PY - 2008/12/1
Y1 - 2008/12/1
N2 - Biogenesis of the mitochondrial respiratory chain enzymes involves the coordinated action of the mitochondrial and nuclear genomes. As a matter of fact, the structural subunits forming these multimeric enzymes are encoded in both genomes. In addition, the assistance of nuclear encoded factors, termed assembly factors, is necessary to allow for the expression of the mitochondrial DNA-encoded subunits and to facilitate their maturation, membrane insertion, and further assembly into the corresponding enzymatic complex. These processes involve transient interactions among the newly synthesized mitochondrial products and specific assembly factors. The identification and characterization of these interactions can be achieved by the method described here, consisting of pulling down tagged versions of the assembly factors immediately after radiolabeling the mitochondrial translation products in isolated mitochondria, and analyzing the radiolabeled pulled-down material.
AB - Biogenesis of the mitochondrial respiratory chain enzymes involves the coordinated action of the mitochondrial and nuclear genomes. As a matter of fact, the structural subunits forming these multimeric enzymes are encoded in both genomes. In addition, the assistance of nuclear encoded factors, termed assembly factors, is necessary to allow for the expression of the mitochondrial DNA-encoded subunits and to facilitate their maturation, membrane insertion, and further assembly into the corresponding enzymatic complex. These processes involve transient interactions among the newly synthesized mitochondrial products and specific assembly factors. The identification and characterization of these interactions can be achieved by the method described here, consisting of pulling down tagged versions of the assembly factors immediately after radiolabeling the mitochondrial translation products in isolated mitochondria, and analyzing the radiolabeled pulled-down material.
KW - GST pull-down
KW - In organello protein synthesis
KW - Mitochondria; mitochondrial biogenesis
KW - Mitochondrial translation
KW - Saccharomyces cerevisiae
UR - http://www.scopus.com/inward/record.url?scp=58149355450&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=58149355450&partnerID=8YFLogxK
U2 - 10.1007/978-1-59745-261-8-9
DO - 10.1007/978-1-59745-261-8-9
M3 - Chapter
C2 - 19066023
SN - 9781588299253
VL - 457
T3 - Methods in Molecular Biology
SP - 125
EP - 139
BT - Methods in Molecular Biology
ER -