Lactating porcine mammary tissue catabolizes branched-chain amino acids for glutamine and aspartate synthesis.
The uptake of branched-chain amino acids (BCAA) from plasma by lactating porcine mammary gland substantially exceeds their output in milk, whereas glutamine output is 125% greater than its uptake from plasma. In this study, we tested the hypothesis that BCAA are catabolized for glutamine synthesis in mammary tissue. Mammary tissue slices from sows on d 28 of lactation were incubated at 37 degrees C for 1 h in Krebs buffer containing 0.5 or 2 mmol/L l-[1-(14)C]- or l-[U-(14)C]-labeled leucine, isoleucine, or valine. Rates of BCAA transport and degradation in mammary tissue were high, with approximately 60% of transaminated BCAA undergoing oxidative decarboxylation and the remainder being released as branched-chain alpha-ketoacids (BCKA). Most ( approximately 70%) of the decarboxylated BCAA were oxidized to CO(2). Rates of net BCAA transamination were similar to rates of glutamate, glutamine, aspartate, asparagine, and alanine synthesis. Consistent with the metabolic data, mammary tissue expressed BCAA aminotransferase (BCAT), BCKA decarboxylase, glutamine synthetase (GS), glutamate-oxaloacetate aminotransferase, glutamate-pyruvate aminotransferase, and asparagine synthetase, but no phosphate-activated glutaminase, activity. Western blot analysis indicated relatively high levels of mitochondrial and cytosolic isoforms of BCAT, as well as BCKA dehydrogenase and GS proteins in mammary tissue. Our results demonstrate that glutamine and aspartate (abundant amino acids in milk protein) were the major nitrogenous products of BCAA catabolism in lactating porcine mammary tissue and provide a biochemical basis to explain an enrichment of glutamine and aspartate in sow milk.
Li
Peng
P
Department of Animal Science and Faculty of Nutrition, Texas A&M University, College Station, TX 77843, USA.
Knabe
Darrell A
DA
Kim
Sung Woo
SW
Lynch
Christopher J
CJ
Hutson
Susan M
SM
Wu
Guoyao
G
D000597:Amino Acids, Branched-Chain / Q000378:metabolism*; D000818:Animals; D001224:Aspartic Acid / Q000096:biosynthesis*; D001692:Biological Transport; D002262:Carboxy-Lyases / Q000378:metabolism; D003653:Decarboxylation; D005260:Female; D005973:Glutamine / Q000096:biosynthesis*; D007651:Keto Acids / Q000378:metabolism; D007774:Lactation / Q000378:metabolism; D008025:Ligases / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism*; D013552:Swine; D014166:Transferases / Q000378:metabolism
D000597:Amino Acids, Branched-Chain; D007651:Keto Acids; D005973:Glutamine; D001224:Aspartic Acid; D014166:Transferases; D002262:Carboxy-Lyases; D008025:Ligases