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Utilization of Supplemental Methionine Sources by Primary Cultures of Chick Hepatocytes

Monsanto Company 800 North Lindbergh Blvd., St. Louis, MO 63167

Utilization of 2-hydroxy-4-(methylthio) butanoic acid (HMB) as a substrate for protein synthesis was studied by using primary cultures of chick liver cells. Cultures were prepared by enzymatic dissociation of livers from week old Hubbard broiler chicks and were maintained for 4 days under nonproliferative conditions. Hepatocyte differentiation was verified by using dexamethasone induction of tyrosine aminotransferase activity. Conversion of [¹⁴C]HMB to L-methionine was shown by chromatographic analysis of hepatocyte protein hydrolysate and incorporation into protein was proven by cycloheximide inhibition of synthesis. When incorporation of HMB was compared to that of DL-methionine (DLM) equimolar quantities of the two sources were found in liver cell protein. These results support, at a cellular level, the conclusion that HMB and DLM are biochemically equivalent sources of methionine for protein synthesis.

KEY WORDS: • methionine • 2-hydroxy-4-(methylthio) butanoic acid • hepatocyte cell culture • methionine hydroxy analogue

Manuscript received 29 April 1983.