lec 29 - Tomography 100 nm 10 nm Shell of Hepatitis B virus...

Info iconThis preview shows pages 1–12. Sign up to view the full content.

View Full Document Right Arrow Icon
Lecture 29: Purification, detection and characterization of proteins II
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Protein sequencing Peptide labeling N-terminal aa cleavage N-terminal aa identification by high-pressure liquid chromatography HPLC) Only sequences for first few amino acids Once a piece of the sequence is known, can be used to search in DNA sequence databases to determine the whole sequence Edman method
Background image of page 2
Mass spectroscopy
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Mass spectroscopy CH 3 OH (MW 32) CH 3 . (MW 15) + . OH (MW 17) ionisation Very precise ( 1 mass unit!) Can determine mass of peptides Can break the peptide bonds and determine mass of fragments differing of one amino acid (each AA has a defined MW) -> deduce sequence
Background image of page 4
Determination of 3D structure X-ray crystallography Cryoelectron microscopy Nuclear magnetic resonance (NMR) spectroscopy
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
X-ray crystallography
Background image of page 6
X-ray crystallography ! Only possible if protein can be crystallized ! -> e.g. almost impossible for most membrane proteins!
Background image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Cryoelectron microscopy
Background image of page 8
Background image of page 9

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 10
Background image of page 11

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 12
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Tomography 100 nm 10 nm Shell of Hepatitis B virus protein dimer NMR In solution -> no need to crystallize-> more “physiological” Only for polypeptides < 200 AAs But can be used to determine structure of domains Molecules exposed to strong magnetic field Measure influence of neighboring atoms on the spin of atoms (generally H and C) Proteomics: comprehensive view of proteins Separation of proteins, e.g. 2D gel electrophoresis cut spot determine sequence (mass spectrometry) Proteomics: examples of applications Comparison of different cell types -> determine cell type-specific Patterns of proteins or protein modifications Comparison of different conditions, treatments (e.g. drugs) Isolation of large complexes/multimolecular assemblies and determination of composition Isolation of organelles -> identification of all components Future: for 50% of genes function still unknown!...
View Full Document

This note was uploaded on 12/07/2010 for the course BIO BIOL 200 taught by Professor Frogatto during the Fall '10 term at McGill.

Page1 / 12

lec 29 - Tomography 100 nm 10 nm Shell of Hepatitis B virus...

This preview shows document pages 1 - 12. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online