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ER bound ribosomes

1. COPI
2. COPII Know the start places and destinations
3. Clathrin
4. Mechanism of the assembly of COPII coated vesicle
Sar1-GDP and Sar1-GTP

5. Mechanism of the assembly of Clathrin coated vesicle


Conception of endocytosis and exocytosis

6. Mechanism of fusion between vesicles and targeted


membrane
Rab-GTP, Snare
Cop II
Inner coat: Sec23 and Sec24
Outer coat: Sec31 and Sec13
GTPase: Sar1
GEF:Sec12

Cop I:
Inner coat: Sec27
Outer coat: Ret1
GTPase: Arf1
GEF:Gea1/Gea2

Clatherin:
Heavy chain
Light chain
Adopter protein: AP2
Dispose waste

Or deliver chemicals to
other cells.

Taking food from out side

Endocytosis is a form of active transport in which a cell transports


molecules into the cells.

Exocytosis: is a form of active transport in which a cell transports molecules


out of the cells
Cargo:
Membrane components and soluble proteins.
Free ribosomes
Import of protein into mitochondria

• Structure of Mit, function


• Signal peptide
• Electron transport chain
• PH of mitochodria (intermembrane space and matrix
space)
• Chaperons
Mitochondrial Organization
•Has its own:
• genome
• tRNA
• rRNA = ribosome
•Encodes some
mitochondrial
proteins
•Most mitochondrial
proteins encoded in
the nucleus
Most mitochondrial
proteins are encoded
in the nucleus

Alberts 14-53
The mitochondria
contains all enzymes
used in the TCA cycle
and oxidative
phosphorylation

Alberts 14-10
The electron transport chain is a set of three proton
pumps that use the energy of NADH to move
protons out of the matrix

Alberts 14-14
Question # 1:
The electron transport chain exports protons out of
the mitochondrial matrix. Therefore:

A. The pH of the Matrix will be lower than that of the


Intermembrane space
B. The pH of the Intermembrane space will be lower
that that of the cytosol (if matrix, this would be
right)
C. The pH of the Intermembrane space will be higher
than that of the Matrix
D. The pH of the Intermembrane space will be the
same as that of the cytosol
E. The pH of the Matrix will be lower than that of the
cytosol
The electron transport chain forms a proton
electrochemical gradient called the Proton Motive Force

Alberts 14-13
Special Protein Translocators Transfer Single Polypeptide
Chains Across and Into Mitochondrial Membrane or Matrix

Alberts 12-23
Mitochondrial Import can be Reconstituted in vitro
Mitochondrial Import can be Reconstituted in vitro

Green = 35 kDa Load onto an


Red = 5 kDa SDS-PAGE

40 kDa ->

20 kDa ->

5 kDa ->
The TOM and TIM23 Complexes Act In Concert
To Transfer Proteins Into The Matrix

Alberts 12-25
The Matrix Targeting Signal (+ charged) moves across
the inner membrane by electrophoresis due to the
Proton Motive Force

Alberts 12-26
Chaperone Pushing and Pulling, and the Proton Motive Force
Contribute To Different steps in Protein Translocation

Alberts 12-26
Mitochondrial HSP70 is required for the
translocation of the peptide into the Matrix

Once in the Matrix the proteins are folded by mHSP-70 and by mHSP-60

Alberts 12-26
Special Protein Translocators Transfer Single Polypeptide
Chains Across and Into Mitochondrial Membrane or Matrix

Alberts 12-23
Special Chaperones and the SAM complex insert Porins into
the outer mitochondrial membrane

The Intermembrane space has its own set of chaperones (not related to HSPs)

Proteins (like Porin) that go through this pathway have a targeting signal different from
MTS but still recognized by the TOM complex.

Alberts 12-27
Multi-pass transmembrane proteins are sorted
to the Inner Mitochondrial Membrane by the
TOM and TIM22 Complexes

This class of proteins includes many metabolite-specific transporters

These proteins also have their own targeting signals

Alberts 12-28
The OXA complex is used to “export” proteins out of the Matrix

OXA can transport proteins that were synthesized within the Matrix OR proteins that
were imported to the Matrix via TOM/TIM23 using an MTS. A separate special signal is
recognized by OXA.
Alberts 12-28
Question # 2:
You are going to measure the import of a protein into
mitochondria in the in vitro assay. You add a drug that
inhibits the electron transport chain. Which of the
following will be a likely effect of adding this drug?

A. The MTS of the protein will not bind to the TOM


receptor
B. Cytosolic Hsp70 will not bind to the protein
C. The MTS of the protein will not pass through the TOM
channel
D. The MTS of the protein will not pass through the TIM
channel
E. The protein will reach the Matrix but will not be cleaved
by the signal peptidase
Import of protein into mitochondria

• Structure of Mit, function


• Signal peptide
• Electron transport chain
• PH of mitochodria (intermembrane space and matrix
space)
• Chaperons

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