Development of Deuterated-Leucine Labeling with Immunoprecipitation to Analyze
Cellular Protein Complex |
| Shufang Liang1,2, Xuejiao Xu2, Haojie Lu2, Pengyuan Yang2 |
| 1State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China |
| 2Department of Chemistry & Institute of Biomedical Science, Fudan University, Shanghai 200433, China |
| Corresponding author: |
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: zizi2006@yahoo.cn |
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| Received August 02, 2008; Accepted September 04, 2008; Published 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, etal. 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. |
| Abstract |
The deuterated-leucine (Leu-d3) labeling is one kind of stable isotope labeling by amino acids in cell culture
(SILAC), which has been widely used to compare and quantify protein relative expression. We expanded an
integrated immunoprecipitation (IP) coupled with SILAC approach (SILAC-IP) to differentiate the specific binding
partners associated with a bait protein in two populations of cells. By this SILAC-IP strategy, the identified
specific-binding proteins were quantified by tracking pairs of Leu-d3 labeled and unlabeled peptides from the
mass spectra, which could differentiate specific-binding proteins from nonspecific partners in high confidence.
We applied SILAC-IP method to differentiate specific-binding proteins associated with 14-3-3â in human hepatocellular
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. |
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