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Figure 15-1 Essential Cell Biology ( Garland Science 2010)

Figure 12-1 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-5 Molecular Biology of the Cell ( Garland Science 2008)

Table 12-1 Molecular Biology of the Cell ( Garland Science 2008)

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Three strategies for sorting of proteins to the appropriate


compartment.
In all cases,
a sorting signal
is incorporated
into the primary
structure of the
nascent
polypeptide.
This sorting signal
is often cleaved
after transport.
Table 12-2 Molecular Biology of the Cell ( Garland Science 2008)

Vesicular Transport System

Protein Sorting Flow Chart


Sorting Signals (AA sequences)
Recognized by Sorting Receptors
that escort protein to the
target compartment

Figure 12-7 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-6 Molecular Biology of the Cell ( Garland Science 2008)

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Signal Sequences and Signal Patches

Table 12-3 Molecular Biology of the Cell ( Garland Science 2008)

Discovery of Signal (Sorting) Sequences


Nuclear Import Pathway
-- P-P-K-K-K-R-K--Internal Signal Sequence
May Be Masked

Figure 15-6 Essential Cell Biology ( Garland Science 2010)

Page 704 Molecular Biology of the Cell ( Garland Science 2008)

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Double Membrane

Nuclear Pore Complex

Inner Membrane is
supported by the
Nuclear Lamina
All macromolecules
that pass into / out of
the nucleus must pass
through a nuclear pore
complex.

Figure 12-8 Molecular Biology of the Cell ( Garland Science 2008)

Nuclear Pore Complex


~ 125,000 kdal
~ 30 different nucleoporins
~ 3000 4000 per nucleus

Figure 12-9 Molecular Biology of the Cell ( Garland Science 2008)

Nuclear Pore Complex (NPC)


Theyre Big ( ~ 80 nm diameter
Small stuff ( < 20,000) gets in by diffusion
Big stuff ( > 60,000) requires escort
~ 500 mcules / sec / NPC
FG repeats

Figure 12-9a Molecular Biology of the Cell ( Garland Science 2008)

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Peptide-Coated Gold Nanospheres

-- P-P-K-K-K-R-L-V---

time after injection

Nuclear Import Signal

In the nucleus

In the cytoplasm

Figure 12-11 Molecular Biology of the Cell ( Garland Science 2008)

Nuclear Import Receptors (NIR) escort large proteins across NPC

Nuclear Import Receptors (NIR) escort large proteins across NPC

Various NIR isoforms recognize different types of proteins


In some cases, adaptor proteins unite cargo with receptor

Various NIR isoforms recognize different types of proteins


Figure 12-13a Molecular Biology of the Cell ( Garland Science 2008)

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In some cases, adaptor proteins unite cargo protein with NIR

Ran-GTPase Cycle controls direction of transport

Figure 12-14 Molecular Biology of the Cell ( Garland Science 2008)

Ran-GTPase Cycle controls direction of transport

Figure 12-14 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-15 Molecular Biology of the Cell ( Garland Science 2008)

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Cargo Protein with NIS


Free NIR
Free NIR binds cargo tightly
Cargo-NIR complex crosses NPC

Ran-GAP hydrolyzes GTP


Ran-GDP dissociates from NIR

Ran-GTP-NIR crosses NPC

NIR binds Ran-GTP tightly


Ran-GTP displaces cargo

Ran-GTP-NIR complex

Ran-GTP decreases
affinity of NIR for cargo

Figure 12-15 Molecular Biology of the Cell ( Garland Science 2008)

Ran-GTP increases
affinity of NER for cargo

Figure 12-15 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-16a Molecular Biology of the Cell ( Garland Science 2008)

Future Dorsal Side

Figure 12-16b Molecular Biology of the Cell ( Garland Science 2008)

Cell Signaling Events can reveal nuclear localization signals


Nuclear Factor of Activated T-cells (NF-AT)

dorsal in cytoplasm

Future Fly

dorsal in nucleus
Future Ventral Side

Figure 12-17 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-18 Molecular Biology of the Cell ( Garland Science 2008)

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Nuclear Lamina on nuclear leaflet of inner nuclear membrane

SEM Image
Figure 12-19 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-20 Molecular Biology of the Cell ( Garland Science 2008)

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Protein Sorting to Mitochondria

Folds to form (+) amphipathic -helix

Some proteins are encoded by


mitochondrial DNA and
biosynthesized in situ.
Most proteins are encoded by
nuclear DNA & made in cytoplasm
Signal = N-terminal, positively
charged amphipathic -helix

Sorting Signal

Proteins are escorted by MIRs


Imported in the unfolded state
Page 713 Molecular Biology of the Cell ( Garland Science 2008)

Four (4) potential destinations

Bound to MIR
Figure 12-22 Molecular Biology of the Cell ( Garland Science 2008)

Mitochondrial Protein
Translocation Complexes

Must cross
2 membranes

Figure 12-21a Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-23 Molecular Biology of the Cell ( Garland Science 2008)

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Translocation to the Mitochondrial Matrix

Energetics of translocation to the Matrix

Powered by (1) ATP hydrolysis


(2) inner membrane electrochemical potential

Figure 12-25 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-26 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-27 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-28 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-28a Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-28b Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-28c Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-28d Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-21 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-29a Molecular Biology of the Cell ( Garland Science 2008)

Protein Sorting to Peroxisomes


Signal = C-terminal SKL
Imported in folded state

Figure 12-29b Molecular Biology of the Cell ( Garland Science 2008)

Page 721 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-30 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-32a Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-32b Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-33 Molecular Biology of the Cell ( Garland Science 2008)

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Endoplasmic Reticulum

Page 723 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-34a Molecular Biology of the Cell ( Garland Science 2008)

Rough Endoplasmic Reticulum


Biosynthesis of membrane & secretory proteins

Figure 12-35 Molecular Biology of the Cell ( Garland Science 2008)

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Smooth Endoplasmic Reticulum


Ca2+ homeostasis / detox (P450) / lipoprotein particles

Figure 12-36b Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-36c Molecular Biology of the Cell ( Garland Science 2008)

Signal Recognition Particle (SRP): an RNA-protein complex

Figure 12-38 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-39b Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-40 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-41a Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-41b Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-42 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-43 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-43c Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-44 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-44a Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-44b,c Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-45 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-46 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-47 Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-47 (part 1 of 2) Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-47 (part 2 of 2) Molecular Biology of the Cell ( Garland Science 2008)

Figure 12-48 Molecular Biology of the Cell ( Garland Science 2008)

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Figure 12-49 Molecular Biology of the Cell ( Garland Science 2008)

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