Preparation of membrane proteins for analysis by two-dimensional gel electrophoresis.
In order to separate hydrophobic membrane proteins, we have developed a novel two-dimensional electrophoresis system. For the iso-electric focusing, agarose was used as a supporting matrix and n-dodecyl-beta-D-maltopyranoside was used as a surfactant. In combination with a previously developed Tris/MES electrophoresis system in the second dimension, distinct spots were reproducibly detected from hydrophobic membrane proteins whose grand average hydropathicity (GRAVY) exceed 0.3. In contrast to the immobilized pH gradient system, c-type heme was also visualized in this system.
Kashino
Yasuhiro
Y
Department of Life Science, University of Hyogo, Ako-gun, Hyogo 678-1297, Japan. kashino@sci.u-hyogo.ac.jp
Harayama
Takeshi
T
Pakrasi
Himadri B
HB
Satoh
Kazuhiko
K
D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D007525:Isoelectric Focusing / Q000379:methods; D008320:Maltose / Q000031:analogs & derivatives / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D045332:Photosystem II Protein Complex / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D012685:Sepharose / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D046939:Synechocystis / Q000378:metabolism
D001426:Bacterial Proteins; D008565:Membrane Proteins; D045332:Photosystem II Protein Complex; C411362:dodecyl maltopyranoside; D008320:Maltose; D012685:Sepharose