BCH3021 Lecture Notes - Lecture 18: Retromer, Protein Targeting, Endosome

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25 May 2018
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Lecture 18 – Protein Sorting in the Trans Golgi Network
Diversity of Cell and Tissue Structure and Function – Reflects complex patterns of
protein distribution
• Many cells are integrated into ordered tissues
• Maintenance of tissue structure and function requires
o Specific cell-cell interactions
o Specific cell protein synthesis and localization
o Cell polarity
• Organelle biogenesis, structure and function require correct protein sorting
• Proteins travel on distinct pathways to arrive at major organelles of cell s
Protein Sorting – general concepts
• Differences between cells in terms of structure and function – differnces in
protein content and localization
• Newly synthesized proteins msut be transported to their site of action (e.g.
cytoplasm, organelle, cell surface) or released into extracellular environment
• Old or defective proteins must be directed to degradative machinery
• Some proteins are itnernalized by cells from extracellular environment
• Tarting of sorting of proteins requires
o A specific signal or address on the protein that directs it to a particular
site
o Specific transport pathways or machinery that recognise the signal and
carry the protein to its final destination
Secretory and Endocytic Pathways are Linked
• Both consist of distinct organelles linked by protein transport pathways
• Proteins are carried in mobile transport vesicles – which travel between
organelles
o Must be sorted or targeted to specific destination
• Golgi pathway – moving things outwards
• Characteristics of each organelle reflect a specific protein content
Protein travels on secretory/endocytic pathways in transport vesicles
• Movement of proteins between organelles can be carried out by different types
of transport vesicles
o COPII vesicle
â–Ş Promotes transfer from ER to golgi
o COPI
â–Ş Golgi to ER
o Clathrin
â–Ş Golgi to lysosome
â–Ş Surface to endosome
o Retromer
â–Ş Endosome to Golgi
o Vesicles move along microtrubules
Golgi is Major Sorting Station of Secretory Pathway
• Once in Golgi, secretory proteins follow one of the several distinct routes or
pathways
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• 1) return to Er via retrieval/recycling pathway
• 2) remain in Golgi as resident protein
• 3) travel to cell surface
• 4) travel to storage organelle for later release
• 5) travel to lysosome
• Proteins are sorted in trans golgi network
o And where they end up is determined by information protein carries
itself
• Proteins are directed into pathways in a sorting organelle – trans Golgi
network
• Specific properties of a protein determine which pathway it follows
o These may include a specific signal motif
Proteins moving between organelles require sorting signals
• Sorting signal – region of a protein required for its correct targeting to a
specific organelle
• Experimentally defined by
o Mutation that disrupts targeting to correct destination
o Transfer of signal to another protein resulting in its re-direction
o Add signal sequence to protein and see transfer of compartment
o Short peptide is necessary and sufficient to direct protein into different
organelle
o Or make point mutation at targeting signal
• Molecular level
o Interacts directly with transport machinery
o Interacts with machinery indirectly through a receptor
Resident ER and Golgi Proteins
• Possession of a signal sequence is necessary and sufficient to direct protein
into ER
• Begin lives in ER then moves through to golgi – requires information on
proteins
• Once a protein has been synthesised in ER – if it doesn’t have any additional
information on it → released to plasma membrane (if has Transmembrane
domain)
o Or if soluble – released from cell
• In absence of additional specific signal, a property folded protein will move
through ER and Golgi and then to cell surface (constitutive pathway or default
pathway)
o Once made in ER, if it doesn’t provide cell with any other signalling
information – it will be released
• Many proteins in ER and Golgi that must remain in these organelles
o Chaperones, oligosaccharyl transferases in ER
o Transferases, sulphatases, phosphotransferases in Golgi
• Protein content of ER and Golgi is maintained by retention (1) and retrieval
(2)
Retrieval Mechanism (being brought back)
• Proteins that have to stay and operate in ER
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