Protein Biology continued . . .

Membrane proteins

Single membrane spanning region

Multiple membrane spanning regions

b-sheet layers

One example of peripheral proteins: Phospholipases

Methods of Protein purification and analysis

Native PolyAcrylamide Gel Electrophoresis (PAGE)

proteins
+

Separation occurs based on charge and on size, but the proteins stay folded in their native conformations

+
Speed of migration will depend on: 1) charge of the protein
2) size of the protein 3) pore size 4) the electric field strength

SDS-PAGE

Denaturing gel includes SDS

2-D electrophoresis

Immunoblotting (aka Western Blotting)

1) Colorimetric development 2) chemiluminescence

3) Radioactivity & autoradiography

Time of flight laser mass spectroscopy

To be continued . . .