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Functionally distinct membrane bound organelles 10 billion proteins of 10,000-20,00 diff kinds Complex delivery system
Compartmentalization of Cells
Membranes
Partition cell Important cellular functions Impermeable to most hydrophobic molecules contain transport proteins to import and export specific molecules Mechanism for importing and incorporating organelle specific proteins that define major organelles
Compartmentalization of Cells
All Eucaryotic Cells Have Same Basic Set of Membrane Bound Organelles
Compartmentalization of Cells
Compartmentalization of Cells
Compartmentalization of Cells
Major Organelles
Nucleus
Cytosol ER Golgi Apparatus Mitochondria and Chloroplast Lysosomes Endosomes Peroxisomes
Compartmentalization of Cells
Occupy 50% cell volume Perform same basic function Vary in size and abundance
Compartmentalization of Cells
Compartmentalization of Cells
Compartmentalization of Cells
Organelles arising from pinching off of pm have interior equivalent to exterior of cell
Compartmentalization of Cells
3 Types of Transport Mechanisms
1. Gated Transport:
gated channels topologically equivalent spaces 2. Transmembrane Transport: protein translocators topologically distinct space 3. Vesicular transport:
Compartmentaliztion of Cells
Families of Intracellular Compartments: 1. nucleus and cytosol 2. organelles in the secretory pathway 3. mitochondria 4. plastid
Compartmentalization of Cells
2 Types of Sorting Signals in Proteins
1. Signal Sequence
continuous sequence of 15-60 aa sometimes removed from finished protein sometimes a part of finished protein
2.
Signal Patch
specific 3d arrangement of atoms on protein surface; aas distant persist in finished protein
Compartmentalization of Cells
Compartmentalization of Cells
Signal Sequences/Patches Direct Proteins to Final Destination
Signal patches direct proteins to: 1. nucleus 2. lysosomes Signal Sequences direct proteins to: 1. ER proteins possess N-terminal signal of 5-10 hydrophobic aa 2. mito proteins have alternating + chg aa w/ hydrophobic aa 3. proxisomal proteins have 3 aa at C-terminus
Compartmentalization of Cells
Sorting signals recognize complementary sorting receptors
Compartmentalization of Cells
Organelles Cannot be Constructed Denovo Organelles reproduced via binary fission Organelle cannot be reconstructed from DNA alone Info in form of one protein that pre-exists in organelle mem is required and passed on from parent to progeny Epigenetic information essential for propogation of cells compartmental organization
Nuclear Envelope
-Outer membrane contiguous w/ER -Inner membrane contains proteins that act as binding sites for chromatin and nuclear lamina
Generally comprised of two short sequences rich in + chged aa lys & arg Can be located anywhere Thought to form loops or patches on protein surface Resident, not cleaved Transport thru lg aqueous pores as opposed to translocator proteins Transports proteins in folded state Energy requiring process
Import Receptors release cargo in nucleus and return to cytosol Export Receptors release cargo in cytoplasm and return to nucleus
Nuclear Export Works like import in reverse Export receptors bind export signals and nucleoporins to guide cargo thru pore Import and export receptors member of same gene family
Controlling rates of import and export determines steady state location phosphorylation/dephosphorylation of adjacent aa may be required for receptor binding Cytosolic anchor or mask proteins block interaction w/ receptors Protein made and stored in inactive form as ER transmembrane protein
Proteins w/ export signals loaded onto RNA during transcription and processing (RNP) Export signals guide RNA out of nucleus thru pores via exportin proteins than bind RNP Export mediated by transient binding to FG repeats Imature mRNAs retained by anchoring to transcription and splicing machinery Proteins disassociate in cytosol and return to nucleus
Nuclear envelop disassembles during mitosis and reassembles when ER wraps around chromosomes and begins to Fuse
TOM- functions across outer membrane TIM- functions across inner membrane OXA- mediates insertion of IM proteins syn w/in mito and helps to insert proteins initially transported into matrix
Complexes contain components that act as receptors and others that form translocation channels
2. Cleavage of N-terminal sequence unmasks 2nd signal used to translocate protein from matrix into or across IM using OXA
3. OXA also used to transport proteins encoded in mito into IM 4. Alternative route bypasses matrix; hydrophobic signal sequence = stop transfer
4.
5. 6.
occur posttranslationally Use separate translocation complexes in ea membrane Translocation occurs at contact sites Requires energy and electrochemical gradient Use amphilpathic N-terminal signal seq that is removed Like the mito a second signal sequence required for translocation into thylakoid mem or space
Peroxisomes
Use O2 and H2O2 to carry out oxidative rxns Remove H from specific organic compounds RH2 + O2 R + H2O2 Catalases use H2O2 to oxidize other substances, particularly in liver and kidney detoxification H2O2 + RH2 R + H2O Beta Oxidation Formation of plasmalogens (abundant class of phospholipids in myelin) Photorespiration and glyoxylate cycle in plants
Peroximsomes in Plants
Site of Photorespiration= glycolate pathway in leaves Called glyoxysomes in seeds where fats converted into sugar
Endoplasmic Reticulum
Occupies >= 50% of cell volume Continuous with nuclear membrane Central to biosyn macromolecules used to construct other organelles Trafficking of proteins to ER lumen, Gogli, lysosome or those to be secreted from cell
ER Central to Protein Synthesis and Trafficking Removes 2 Types of Proteins from Cytosol: 1. 2. transmembrane proteins partly translocated across ER embedded in it water soluble proteins translocated into lumen
Hydrophobic signal sequence of diff sequence and shape SRP lg hydrophobic pocket lined by Met having unbranched flexible side chains Binding of SRP causes pause in protein synthesis allowing time for SRP-ribosome complex to bind to SRP receptor
Protein to be imported passes through an aqueous pore in the translocator that is a dynamic structure
Sec61 protein translocator
Signal sequence triggers opening of pore Translocator pore closes when ribo not present
Some proteins are imported in to ER by a posttranslational mechanism Proteins released into cytoplasm Binding of chaperone proteins prevents them from folding Translocation occurs w/out ribo sealing pore Mechanism whereby protein moves through pore unkwn
Two signaling functions: 1) directs protein to ER membrane 2) serves as start transfer signal to open pore Signal peptidase removes terminal ER signal sequence upon release of protein into the lumen
N-terminal signal sequence initiates translocation and additional hydrophobic stop sequence anchors protein in membrane Signal sequence is internal and remains in lipid bilayer after release from translocator Internal signal sequence in opposite orientation Orientation of start-transfer sequence governed by distribution of nearby chg aa
2.
3.
4.
Combinations of start- and stop-transfer signals determine topology Whether hydrophobic signal sequence is a start- or stop-transfer sequence depends upon its location in polypeptide chain All copies of same polypeptide have same orientation
ER resident proteins contain an ER retention signal of 4 specific aa at Cterminus PDI protein disulfide isomerase oxidizes free SH grps on cysteines to from disulfide bonds S-S allowing proteins to refold BiP chaperone proteins, pulls proteins posttranslationally into ER thru translocator and assists w/ protein folding
Most soluble and transmembrane proteins made in ER are glycolsylated by addition of an oligosaccharide to Asn Precursor oligosaccharide linked to dolichol lipid in ER mem, in high energy state Transfer by oligosaccharyl transferase occurs almost as soon as polypeptide enters lumen
Retrotranslocation
Improperly folded ER proteins are exported and degraded in cytosol Misfolded proteins in ER activate an Unfolded Protein Response to increase transcription of ER chaperones and degradative enzymes
ER synthesizes nearly all major classes of lipids Phospholipid synthesis occurs on cytoplasmic face by enzymes in mem Acyl transferases add two FA to glycerol phosphate producing phosphatidic acid Later steps determine head group
Scramblase phospholipid translocator equilibrates phospholipids distribution Flipasses of PM responsible for asymmetric distribution of phospholipids