Development of Deuterated-Leucine Labeling with Immunoprecipitation to Analyze Cellular Protein Complex
- *Corresponding Author:
- Dr. Shufang Liang.
State Key Laboratory of Biotherapy
West China Hospital, Sichuan University
1# Keyuan Road 4, Gaopeng Street
Chengdu 610041, China
Fax : +86 28 85164060
E-mail: [email protected]
Received Date: August 02, 2008; Accepted Date: September 04, 2008; Published Date: September 13, 2008
Citation: Shufang L , Xuejiao X , Haojie L , Pengyuan Y (2008) Development of Deuterated-Leucine Labeling with Immunoprecipitation to Analyze Cellular Protein Complex. J Proteomics Bioinform 1:293-301. doi:10.4172/jpb.1000037
Copyright: © 2008 Shufang L, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
The deuterated-leucine (Leu-d 3 ) labeling is one kind of stable isotope labeling by amino acids in cell culture (SILAC), which has been widely used to compar e and quantify pr otein r elative expr ession. W e expanded an integrated immunoprecipitation (IP) coupled with SILAC approach (SILAC-IP) to differentiate the specific bind- ing par tners associated with a bait pr otein in two populations of cells. By this SILAC-IP strategy , the identified specific-binding proteins were quantified by tracking pairs of Leu-d 3 labeled and unlabeled peptides from the mass spectra, which could differentiate specific-binding proteins from nonspecific partners in high confidence. W e applied SILAC-IP method to dif fer entiate specific-binding pr oteins associated with 14-3-3â in human hepato- cellular carcinoma cell line QGY7703 between those in the liver cell line QSG7701. The proteins including HSP86, SKB1hs, GADPH and MEP50 were identified to associate with 14-3-3â in QGY7703 with high binding level than in QSG7701 cells.