Fuctional Proteomics On Acute Phase Response Of Loach To Trauma

Background:Innate immunity system provide the body with its first line of protection against external injury and such a rapid,non-specific mechanism can almost be foundall through the living kingdom. Both cells and secreted molecules of innate immunity are dynamic in regard to their readiness status and the process of cell activation during the adaptive alteration of cell physiology at the initiation stage of innate immunity is addressed as acute phase response (APR). It involves a large number of functional proteins (APP) and functions in a variety of defense-related activities such as limiting the dispersal of infectious agents,repair of tissue damage,killingof microbes and restoration of healthy state. Although history has witnessed fruitful achievement in the field of mammal APR,especially in the field of humanclinical medicine,the fact that innate defense system plays more important rolein the self-protection of lower animals thus they have robust APR system is worthmuch attention as well. Development of proteomics provides rapid,high sensitive,high throughput approach for study of such a complex biological problems as APR froma comprehensive point of view which overcomes the defects of classical targetedprotein analysis including low efficiency and failure in identifying new APPcandidate.Objective:Study the proteome profiles of serum and skin proteins of loach stimulatedwith trauma in comparison to control group. Identify APP candidates and search forfunctional proteins of special research value.Methods and Results:Proteomics approaches involve three critical techniques:twodimensional electrophoresis,biological mass spectrometry and bioinformatics. In2-DE,we applied traditional IEF in the first dimension and Tris-tricine system inthe second dimension and acquired protein profiles of serum and skin samples. In the identification of 10 proteins of serum and 16 proteins of skin,we acquired peptide mass of fingerprint (PMF) maps for all targets by means of matrix-assistedlaser desorption-ionization time of flight mass spectrometry (MALDI-TOF-MS) andpartial aminio acid sequences for 9 proteins by means of electrospray ionizationms/ms(liSI-MS/MS). 22 proteins were identified through matching the above informal ion with Swissl'rot database,among which 3 proteins of serum as well as 5 proteins of skin were reported APPs and other proteins were latent APP candidates. We also found an angiostatin-like protein and the detection of its activity on chick embryo chorioallantoic membrane model (CAM) shows a sign of inhibition of the positive effect of lung cancer cell culture supernatant on CAM. Conclusion:Development of proteomics brings out highly efficient approach for APR research,which overcomes the drawbacks of traditional methods including low efficiency and difficulty in detecting new APR-related proteins and the intrinsic false positive reaction of recently developed SSH technology.