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IUBMB
Review Article
Translocation of Proteins into Mitochondria
Nicholas J. Hoogenraad and Michael T. Ryan
Department of Biochemistry, La Trobe University, 3086 Melbourne, Australia
Summary
The translocase of the outer mitochondrial membrane (TOM)
is composed of receptors, a channel protein, and its modulators
that function together to import proteins into mitochondria. Although the import pathway of proteins directed to the mitochondrial matrix has been well characterized, recent studies into the
import pathway taken by proteins into the other submitochondrial
compartments have broadened our understanding into the way the
TOM machinery recognizes, interacts, and translocates proteins.
IUBMB Life, 51: 345 350, 2001
Keywords
INTRODUCTION
Mitochondria are essential for the viability of eukaryotic
cells. Whereas they are the main source of respiratory energy,
cells in culture can survive the complete depletion of mitochondrial DNA, which encodes 13 polypeptides of the oxidative phosphorylation machinery. This suggests that mitochondria are involved in essential activities besides the production
of ATP. In fact, mitochondria are the major source of damaging reactive oxygen species and contain key regulators of programmed cell death (apoptosis). There has also been an emerging realization that mitochondrial dysfunction is at the center
of medical conditions ranging from early onset inborn errors of
metabolism to later onset neuromuscular disorders and a wide
range of unexpected phenotypes including Parkinsons Disease
and diabetes (1).
Mitochondria grow and divide by a process of accretion of
proteins encoded in the nucleus and mitochondria, followed by
binary ssion. The 1,000 nuclear encoded proteins need to be
imported into mitochondria (2, 3). Proteins are rst synthesized
in the cytosol on free ribosomes as precursors with signals that
direct them to one of the four mitochondrial subcompartments
the outer membrane, intermembrane space (IMS), inner
Received 28 June 2001; accepted 13 August 2001.
Address correspondence to Nicholas J. Hoogenraad. Fax: 61-39479-2467. E-mail: N.Hoogenraad@latrobe.edu.au
membrane, and matrix. A typical matrix-targeted precursor contains an N-terminal presequence 15 to 40 amino acids long that
is rich in basic and hydroxylated amino-acids and has the potential to form an amphipathic -helix. Whereas some inner
membrane and intermembrane space proteins also possess such
a presequence, others do not and are like all outer-membrane
proteins in that they contain their often cryptic mitochondrial
targeting signal(s) within the sequence of the mature protein.
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346
4-Color
art
Figure 1. Import of proteins into mitochondria. Typical matrix-targeted precursors (blue) are targeted to the mitochondrial outer
membrane via positive N-terminal targeting signals, where they are mainly recognized by the receptor Tom20. Some precursors
destined for the inner membrane, such as AAC (red), contain internal targeting signals and bind to the receptor Tom70. Overlapping
speci cities between the receptors for precursors may be observed. The precursors are transferred to other members of the TOM
machinery (green) before their translocation across the Tom40 channel. There, the pathways diverge. Matrix-targeted precursors
engage with the Tim23 complex and are translocated into the matrix in a membrane potential dependent manner. Some of these
precursors contain stop-transfer signals and are instead released into the inner membrane. If proteolytically processed by an IMS
protease, they are released into the IMS. AAC associates with Tim9 and Tim10 in the IMS before binding to the Tim22 complex
and inserting into the inner membrane in a membrane potential dependent manner.
motifs and to have the ability to distinguish those hydropho bic membrane proteins for insertion into the outer membrane
from those that must bypass the outer membrane and be targeted
and inserted into the inner membrane. In addition, mechanisms
must be in place to ensure translocation occurs in a unidirectional manner. Whereas precursor-binding sites are present on
the intermembrane space (or trans-) side of the outer membrane,
precursors must still rst insert, travel along, and then exit the
TOM channel before such interactions can take place.
Tom20, Tom6, and Tom7 (5). A Tom5 homologue was not identi ed but may be present in low amounts. In yeast, a 400-kDa
TOM complex has been readily observed by Blue Native PAGE
and contains Tom40, Tom22, Tom7, Tom6, and Tom5 (6). Under
gentle solubilization conditions, Tom20 can also be found associated with this complex (7 ). The mammalian TOM complex
contains homologous subunits and migrates slightly faster than
the yeast complex on Blue Native PAGE (8).
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348
349
tined for different mitochondrial subcompartments. These studies could also determine the functional relevance of multiple
channels within a single complex. Perhaps some asymmetry exists between the different channels of a single complex that enable the import machinery to discriminate between precursors
that must insert into the outer membrane and those that must
translocate across the outer membrane.
ACKNOWLEDGMENTS
We apologize to the many investigators whose work could
not be cited due to space limitations. We thank A. Johnston
for critically reading the manuscript. This work is supported by
grants from the Australian Research Council.
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