Premium
Regulation of protein turnover in porcine intestinal cells by L‐glutamine (Gln)
Author(s) -
Xi Pengbin,
Jiang Zongyong,
Dai Zhaolai,
Li Xilong,
Yao Kang,
Wang Junjun,
Wu Guoyao
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.740.7
Subject(s) - glutamine , microbiology and biotechnology , cell culture , protein biosynthesis , chemistry , biochemistry , biology , amino acid , genetics
Intestinal porcine epithelial cells (IPEC‐1) from newborn pigs were seeded in 6‐well culture plates with DMEM‐F12 containing 5% FBS, 1% antibiotics and 5μg/ml insulin. For determining protein degradation, after 16‐h culture, cells were cultured for 3 days in DMEM‐F12 containing 0.5 mM Gln. Beginning on day 4, cells were cultured for 24 h in DMEM containing 0.5 mM Gln and 0.1 mM L‐phenylalanine (Phe) plus L‐[ 3 H]Phe. After 24‐h culture, the medium was removed and cells were washed 3 times with PBS. The cells were then cultured for 3 h in 2 ml DMEM containing 1 mM Phe and 0.03, 0.5 and 2 mM Gln. At the end of 3‐h culture, medium and cells were analyzed for 3 H‐Phe. For determining protein synthesis, after 16‐h culture, the cells were cultured for 3 days in DMEM‐F12 containing 0.5 mM Gln. On day 4, cells were cultured for 3 h in 2 ml DMEM containing 1 mM Phe plus 0.1 μCi L‐[ring‐2,4‐ 3 H]Phe, 0.03, 0.5 and 2 mM Gln. At the end of 3‐h culture, cells were collected to determine protein‐bound 3 H‐Phe. Compared with 0.03 mM Gln, addition of 0.5 and 2 mM Gln to culture medium dose‐dependently decreased (P < 0.05) protein degradation in IPEC‐cells by 19% and 23%, respectively, while increasing protein synthesis by 28% and 36%, respectively, in IPEC‐1 cells. Likewise, 0.5 and 2 mM Gln enhanced the growth of IPEC‐1 cells. These results indicate that Gln stimulates net protein synthesis in intestinal cells. Supported by grants from USDA and NSFC.