Intracellular Compartments and Protein Sorting Haixu Tang School of Informatics The major intracellular compartments of an animal cell Relative Volumes Occupied by the Major Intracellular Compartments INTRACELLULAR COMPARTMENT PERCENTAGE OF TOTAL CELL VOLUME Cytosol 54 Mitochondria 22 Rough ER cisternae 9 Smooth ER cisternae plus Golgi cisternae 6 Nucleus 6 Peroxisomes 1 Lysosomes 1 Endosomes 1 An electron micrograph Development of plastids Hypothetical schemes for the evolutionary origins of some organelles Four distinct families 1) the nucleus and the cytosol, which communicate through nuclear pore complexes and are thus topologically continuous (although functionally distinct); 2) all organelles that function in the secretory and endocytic pathways, including the ER, Golgi apparatus, endosomes, lysosomes, the numerous classes of transport intermediates such as transport vesicles, and possibly peroxisomes; 3) the mitochondria; 4) the plastids (in plants only). Secretory vs. endocytic pathways Protein traffic Protein traffic • Gated transport • Transmembrane transport • Vesicular transport – membrane-enclosed transport intermediates Sorting sequences Some sorting sequences Prediction of protein sorting • Psort web server: http://psort.nibb.ac.jp/ – prediction of protein localization sites in cells from their primary amino acid sequence Construction of Membrane-enclosed Organelles Require Information in the Organelle Itself • The information required to construct a membraneenclosed organelle does not reside exclusively in the DNA that specifies the organelle's proteins. Epigenetic information in the form of at least one distinct protein that preexists in the organelle membrane is also required, and this information is passed from parent cell to progeny cell in the form of the organelle itself. Presumably, such information is essential for the propagation of the cell's compartmental organization, just as the information in DNA is essential for the propagation of the cell's nucleotide and amino acid sequences. Nuclear pore complexes Nuclear Envelope Nuclear lamina • Consists of "intermediate filaments", 30-100 nm thick. • These intermediate filaments are polymers of lamin, ranging from 60-75 kD. • A-type lamins are inside, next to nucleoplasm; B-type lamins are near the nuclear membrane (inner). They may bind to integral proteins inside that membrane. • The lamins may be involved in the functional organization of the nucleus. Nuclear localization signals (NLSs) Protein import through nuclear pores Possible paths for free diffusion through the nuclear pore complex Nuclear Import / Export Receptors The control of nuclear import during T-cell activation The breakdown and re-formation of the nuclear envelope during mitosis The subcompartments of mitochondria and chloroplasts A signal sequence for mitochondrial protein import Protein translocators in the mitochondrial membra Protein translocation depends on the temperature Protein import by mitochondria Energy required Two plausible models of how mitochondrial hsp70 could drive protein import Protein import from the cytosol into the inner mitochondrial membrane or intermembrane space Translocation of a precursor protein into the thylakoid space of chloroplasts The Endoplasmic Reticulum Free and membrane-bound ribosomes The signal hypothesis The signal-recognition particle (SRP) SRP direct ribosomes to the ER membrane Protein translocation Single-pass transmembrane protein Multipass membrane protein rhodopsin Protein glycosylation in the rough ER The export and degradation of misfolded ER proteins The unfolded protein response in yeast Phospholipid exchange proteins