Differential Proteins of Liver Revealed with a Combined Technology of both Differential Centrifugation and Proteomics in Paralichthys Olivaceus Under Stress of Cadmium Salt

Abstract

Both total proteome and differential proteins were effectively extracted, separated and selected by a combined approach of both differential centrifugation and 2D-PAGE in liver of Paralichtys olivaceus (POL) under the stress of cadmium chloride as an artificial pollution source. Approximately 800 spots for extraction of whole protein separated with 2D-PAGE were obtained by direct lysis in the POL. In addition, approximately 11 differential proteins in POL were also obtained under the stress of cadmium chloride. The differential centrifugal were used to prepare three sedimentation and a plasmolysis proteins, called POL component I, POL component II, POL component III, and POL component IV (plasmolysis protein), respectively. Total protein spots for each gel were calculated to have 380, 550, 500, and 850, respectively, approximately 2280 spots in sum, while total spots are much higher than those by direct lysis approach. Using the comparison method, approximately 54 differential proteins in POL were obtained by a combined technology of both differential and two dimensional polyacylaminde gel electrophoresis(2 D-PAGE) methods under the stress of cadmium chloride. In addition, these differential proteins can be further identified by peptide mass fingerprint (PMF). Here, these combined techniques can be effectively used to extract, separate and identify the whole proteins and the differential proteins including protein markers in the biological tissue.