Introduction •They are found on the outer membrane of mitochondria and chloroplast (Joreset al.2018) •Mitochondrialβ-barrel proteins are found in the nucleus. •They are translated by the cytosolic ribosomes •They perform some important functions like the transport of small molecules including ions and nucleic acids.
Introduction •They have important role in the transport of proteins that are synthesized in the cytosol and in the exchange of lipids •The family of theATP-dependent chaperones of the heat shock protein 70 (Hsp70) help in the scaffolding of the newly synthesized proteins •They can also open and can also cause the disaggregation of the misfolded proteins (Joreset al.2018) •Hsp40 is a cochaperone that helps in the formation of complex between the Hsp70 and the other client proteins.
Method •The use of the Hsp chaperones has been proved by the use of the yeast strains .The tom70/71Δ strain was prepared by successive knockouts. Then to check the products In vitro translation and import of radiolabeled proteins was carried out. •After this the product was centrifuged to remove ribosomes. •Then the import of the porin protein was analyzed and the human Tom40 was analyzed by proteinase K (PK) treatment (Jores et al. 2018).
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Methods •Then after this thePull-down of in vitro translated proteins was performed •The proteins that were translated in the yeast extract were pulled down. •After that the protein was analyzed bySDS-PAGE followed by Western blotting or mass spectrometry (Jores et al. 2018).
Method •The next method is theUV-induced cross-linking. •The yeast cells were transformed with plasmid and then irradiated with UV lamp and the remaining sample was kept in dark •After the treatment the samples were subjected to analysis either by mass spectrometry or SDS-PAGE and Western Blotting (Jores et al. 2018).