Biomedical Engineering Reference
In-Depth Information
A second novel spike for translocation research came from
the identifi cation of entire proteomes of individual cellular
compartments. For example, the analysis of the mitochondrial pro-
teome [
66
] initiated a boom in the search for novel components,
which signifi cantly improved our understanding of the composition
and function of mitochondrial protein translocases. Comprehensive
proteomic studies have been recently published for most compart-
ments of animal, yeast, or plant cells [
67
-
76
].
The impact of systems biology on our current understanding
of how the cell works is invaluable. Nevertheless, without the
detailed analysis of the molecular structure and function of indi-
vidual components all “OMICs” approaches would only provide
inventory lists and interaction networks. Therefore, a thorough
characterization of specifi c factors identifi ed is more important
than ever. In addition to classical biochemical and cell biological
strategies which are outlined in many chapters of this topic, insights
into the structure of proteins have shown to be very helpful.
An excellent example is the solution of the structure of the Sec
translocase by the group of Tom Rapoport [
77
] which signifi cantly
inspired the entire fi eld of protein translocation. In addition to
crystallization studies the use of cryo-electron microscopy and
modern spectroscopic strategies allowed stimulating insights into the
structure of translocation complexes (for examples
see
ref.
78
-
80
).
Also the vesicle budding fi eld has greatly benefi ted from recently
published structures like those of the coat components [
30
,
32
,
62
,
81
,
82
]. These studies provided the fi rst evidence for a direct
interaction of SNARE proteins with a subunit of the COPII coat.
Two major fi ndings revolutionized the way cell biologist con-
duct their experiments today: the discovery of green fl uorescence
protein as a localization tool and the applicability of siRNA knock-
downs and CRISP-Cas9 knockouts in different experimental sys-
tems [
83
-
85
]. These technologies combined with high-throughput
automated microscopy have started and will continue to increase
the knowledge on the communication between and the dynamics
of intracellular organelles tremendously. Moreover, super-resolution
microscopy and now the reliable use of correlative microscopy will
further enhance understanding of different intracellular transport
Baumann et al., Chapter
24
; Mironov and Beznoussenko, Chapter
23
;
Hirschmann et al., Chapter
25
)
. All the puzzle pieces contributed
by the different strategies of systems biology, biochemistry, cell
biology, genetics, and structural biology promise—when combined
into one big picture—an exciting full view on the processes by
which eukaryotic cells achieve the intracellular traffi cking of their
constituents.
