Does glutamine act as a substrate for transamination reactions in the liver of fed and fasted sheep?
The present study investigated the relative importance of glutamine as a transamination source in the ovine liver by examination of the labelling of amino acids (AA) in the hepatic free pool, mixed liver and plasma proteins of fed and fasted sheep, following infusion of isotopically-labelled glutamine. In a cross-over design four sheep were either fasted for 3 d or fed to maintenance and finally euthanased. At each intake, the sheep were infused for 6 h with [2-15N]glutamine (150 micromol/h) and samples of total plasma protein isolated. Following the terminal infusion, liver tissue total proteins were prepared and hydrolysed and 15N-enrichments in seventeen AA were determined by GC-combustion-isotope-ratio mass spectrometry. All AA were enriched (relative to natural abundance) except lysine and threonine, with the lowest enrichments in phenylalanine and histidine. There was no effect of the fed v. fasted state, except for leucine and isoleucine in liver protein Enrichments in liver protein were greater than in plasma protein except proline) and probably reflect the faster turnover rate of hepatic constitutive proteins compared with export proteins. Amination to methionine was greater than that to phenylalanine suggesting a mechanism for preferentially protecting the former. This factor could be important for ruminant production, as methionine is often considered to be the first limiting AA for animals offered certain silages and conserved forages. Enrichments in all AA (except for glutamine, alanine and aspartate) were less than that for glutamate and thus transaminations may have occurred with glutamine directly or via glutamate, following the action of hepatic glutaminase.
Hoskin
S O
SO
Rowett Research Institute, Bucksburn, Aberdeen AB21 9SB, UK.
Gavet
S
S
Milne
E
E
Lobley
G E
GE
D000704:Analysis of Variance; D000818:Animals; D018592:Cross-Over Studies; D005215:Fasting; D008401:Gas Chromatography-Mass Spectrometry; D005972:Glutaminase / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D009587:Nitrogen Isotopes / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D012756:Sheep / Q000378:metabolism*
D009587:Nitrogen Isotopes; D011506:Proteins; D005973:Glutamine; D010649:Phenylalanine; D008715:Methionine; D005972:Glutaminase