Two-dimensional analysis of recombinant E. coli proteins using capillary isoelectric focusing electrospray ionization mass spectrometry.
On-line combination of capillary isoelectric focusing with electrospray ionization mass spectrometry is applied for a two-dimensional analysis of Escherichia coli proteins. The proteins are focused and cathodically mobilized in a polyacrylamide coated capillary. At the end of the capillary, various protein zones are analyzed by mass spectrometry coupled through an electrospray interface. Comparisons with silver-stained two-dimensional gel electrophoresis are made with regard to mass determination, resolution, speed, and sensitivity. Direct identification of a recombinant fusion protein of glutathione S-transferase and striped bass growth hormone is achieved without any prior protein isolation procedures.
Tang
W
W
Department of Chemistry, USDOE, Iowa State University, Ames 50011, USA.
Harrata
A K
AK
Lee
C S
CS
D000665:Ampholyte Mixtures; D000818:Animals; D001492:Bass; D003001:Cloning, Molecular; D019075:Electrophoresis, Capillary / Q000379:methods*; D004926:Escherichia coli; D005822:Genetic Vectors; D005982:Glutathione Transferase; D013006:Growth Hormone / Q000032:analysis* / Q000235:genetics; D007525:Isoelectric Focusing / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis*
D000665:Ampholyte Mixtures; D011993:Recombinant Fusion Proteins; D013006:Growth Hormone; D005982:Glutathione Transferase