Name: Translocation of Proteins into the Mitochondria
Uploaded: 2023-04-30T09:41:52
Description: Learn by watching this video about Translocation of Proteins into the Mitochondria at JoVE.com

Peptides carrying presequences targeted to the matrix are threaded through the TOM/TIM complex. Mitochondrial Hsp70 binds the peptide as it exits the TIM channel and moves it into the matrix, where the presequence is cleaved by matrix proteases.
Proteins targeted to the intermembrane space carry an additional hydrophobic signal sequence that stops translocation across the TIM complex. Signal peptidases cleave the hydrophobic segment and release the active protein in the intermembrane space.
Alternatively, mitochondrial intermembrane space assembly 40 or Mia40 protein imports intermembrane space proteins lacking the presequences.
Oxidized Mia40 forms a transient disulfide bond with the thiols on the incoming polypeptide and pulls the nascent peptide through the TOM channel. Once the entire peptide is threaded through, Mia40 is reduced.
Some inner membrane proteins with stop-transfer sequences are arrested and inserted into the membrane. Others are first processed by signal peptidases, and then recognized by an OXA&#160; translocase, which embeds them in the membrane.

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	<li>Diana Stojanovski et al., The MIA pathway: A tight bond between protein transport and oxidative folding in mitochondria. Biochimica et Biophysica Acta 1823 (2012) 1142-1150.</li>
	<li>Nils Wiedemann and Nikolaus Pfanner. Mitochondrial Machineries for Protein Import and Assembly. Annu. Rev. Biochem. 2017. 86:685&#8211;714.</li>
	<li>Akio Ito. Mitochondrial Processing Peptidase: Multiple-Site Recognition of Precursor Proteins. Biochemical and Biophysical Research Communications 265, 611&#8211;616 (1999)</li>
</ol>

translocation of proteins into the mitochondria

Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast, alpha-helical membrane-anchored proteins are translocated by the TOM complex and inserted by the mitochondrial import complex.
Sorting of intermembrane space proteins:
Intermembrane space proteins are sorted via the mitochondrial intermembrane space import and assembly machinery or (MIA) pathway. MIA40 is an oxidoreductase of the intermembrane space (IMS) that recognizes and binds specific cysteine residues upstream or downstream of the hydrophobic intermembrane space sorting sequence called the MISS/ITS signal. MIA40 contains a redox-active CPC motif that facilitates thiol-disulfide exchange with the cysteine residues of the incoming precursor. Dimeric flavin-dependent oxidoreductase called Erv1 cooperates with MIA40 in oxidizing the precursor cysteine residues, thereby folding and transporting the oxidized precursor into the IMS. Erv1 then transfers electrons to the reduced CPC motif of MIA40 via cytochrome C and cytochrome C oxidase or respiratory chain IV of the electron transport chain. Oxidized Mia40 gets ready for another round of IMS precursor import.
Sorting of inner membrane and matrix proteins:
Inner membrane proteins with cleavable N terminal presequences are translocated through the presequence translocase TIM23 assisted by the ATP-dependent presequence translocase associated motor (PAM). In contrast, inner membrane proteins with internal import signals called carrier signatures are translocated via the carrier translocase TIM22.&#160; Translocation across TIM23 or TIM22 to the inner membrane can follow two distinct routes of sorting: the conservative and stop transport pathways. In the stop-transfer pathway, the movement of precursors with internal hydrophobic patches gets blocked across the TIM22/23 channel, which gets laterally released onto the inner membrane. In the conservative pathway, precursors are first translocated to the matrix and then assembled into the inner membrane by the Oxa protein. However, precursors targeted to the matrix are cleaved by matrix processing peptidases. Some precursors are further processed by mitochondrial intermediate peptidase or inner membrane protease I depending on additional presequence cleavage sites. Processed matrix proteins are then released into the matrix to be folded by chaperonins using ATP hydrolysis energy.

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