march 17 notes - March 17 DVD Lectures Rocks Protein...

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March 17 DVD Lectures Rocks Protein synthesis and trafficking Begins on polyribosomes 2 paths o ER If protein has signal sequence recognized by SRP If synthesized here, have very specific functions o Polyribosomes Determined by targeting sequence Protein Translocation Look at previous notes Translocon o How do we know Translocon Exists? Experiments Simon and Blobel o 15 years old o Realized TEM was wrong o Process Took Cells Make rough microsomes with nascent proteins Developed 2 chamber system separated by artificial membrane Took rough microsomes and fused with this membrane Not vesicle, but a sheet n middle of membrane that has ribosomes on one side and nascent protein going through the protein o Why? Wanted to see the hole in a different way. Measure transmembrane potential Took Ohms Law, took voltage across membrane Voltage should stay the same o If Resistance, then there is a change in Porosity; there is a hole that was not there before. Add Puromycin Works by extruding microsomes Ribosomes keep the pores Decrease in resistance
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o There is a hole present in the membrane Add Sodium Chloride, 400 millimolar Ribosomes disassociate Resistance measured, went up o Now holes are closed Conclusion There must be some complex pores present Presence of ribosomes keeps pores open Photoaffinity label o Translocon with Ribosome o Synthesized so that once in translocation, it can be covalently bonded If Radioactive, you can immunoprecipitate or Gel or
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This note was uploaded on 04/13/2009 for the course BIOL 311 taught by Professor Vanbuskirk during the Spring '08 term at Binghamton University.

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march 17 notes - March 17 DVD Lectures Rocks Protein...

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