Screening The Proteins Interacting With Intelectin-1 In Crohn Disease By Co-immunoprecipitation And MALDL-TOF/TOF-MS

Crohn disease(CD)is a non-specific inflammatory bowel disease with complicated mechanisms and uncertain pathogenesis.CD could occur in any part of the digestive tract,especially the ileocecal area,sometimes accompanied with parenteral manifestations.A variety of pathogenic factors(such as infection,environment,autoimmune disorders and genetic factors)may contribute to CD.In the past 20 years,the incidence of Crohn’s disease increased year by year with the development of industrialized urban processes in China.Because of its recurrent and delayed healing,CD seriously reduce the quality of patients’ life and should cause clinicians attach great importance.The damage of intestinal mucosal barrier is the key point in the research of CD.Intelectin-1(ITIN1)is a secretory glycoprotein mainly expressed in lung,heart,small intestine and colon,which can participate in intestinal immune defensing against microbes,insulin-stimulated glucose uptaking,chronic obstructive pulmonary disease and asthma,etc..Through previous study,we found that Intelectin-1 protein wes highly expressed in the Crohn’s patients’ involved intestinal mucosa,compared to the normal intestinal mucosa.Suggesting that the Intelectin-1 protein may play a important role in the pathogenesis of Crohn’s disease.In this study,protein complex of Intelectin-1 were purified and identified.The Intelectin-1 complex was purified from Crohn’s patients’ involved intestinal mucosa via the method of coimmunoprecipitation with specific antibody.Stained by Coomassiebrilliant blue after SDS-PAGE electrophoresis.Then cut off the band different from the control and identified the complex by Mass Spectrometry.In all,4 Intelectin-1related proteins were identified,including:TRAF3、ATPase、HSP90、ZNF.The validation of Intelectin-1-interaction proteins with immunoprecipitation and Western blot.And identified the interaction among TRAF3 protein and Intelectin-1 in diseased tissue of Crohn’s disease patient.It is said these proteins could play a major role in the development of CD,presumably,Intelectin-1 may through their interactions to the effect.