Fractionation of soluble proteins in Escherichia coli using DEAE-, SP-, and phenyl sepharose chromatographies.

In an effort to simplify a complex mixture of soluble proteins from Escherichia coli, methods to fractionate the samples prior to two-dimensional (2D) gel electrophoresis were developed. These methods involve the use of DEAE-Sepharose, SP-Sepharose, and phenyl Sepharose chromatographic columns and the fractionation of the protein mixtures based on differential anionic, cationic, and hydrophobic properties of the proteins, respectively. Fractionation of the soluble proteins from an E. coli extract with DEAE-Sepharose resulted in a threefold increase in the number of detectable 2D gel spots. These gel spots were amenable to protein identification by using in-gel trypsin digestions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and peptide mass fingerprinting. Significantly, the DEAE-Sepharose column fractionation effectively partitioned the soluble proteins from the cell extracts. Similarly, an SP-Sepharose column was used to fractionate the soluble proteins from E. coli and resulted in over a twofold increase in the number of detectable gel spots. Lastly, fractionation of the cell extract with the phenyl Sepharose column resulted in a threefold increase in the number of detectable 2D gel spots. This work describes an easy, inexpensive way to fractionate the soluble proteins in E. coli and a way to better profile the E. coli proteome.     

Alewife,rainbow smelt and native fishes in Lake Michigan: competition or predation?

Synopsis Several native fish species in Lake Michigan became rare or locally extinct during the increase of rainbow smelt (1930s) and alewife (1950s–1960s). These particular native species have pelagic eggs or larvae which were large relative to the zooplankton and which co-occurred with feeding alewife, smelt or both. Alewife, smelt and most of the other planktivores in the lake probably consumed eggs, and at least alewife and smelt are known to consume larvae. For each species, I comment on the probable role of predation on early life history stages in their decline. Several species which currently co-exist with alewife and smelt have shown large scale declines during the increase of the exotics. Recruitment declines were often dramatic and the probability of predation on these species during their early life history is evaluated.