TY - JOUR
T1 - In vitro amino acid incorporation by the post-mitochondrial supernatant from rat liver
AU - Richardson, Arlan
AU - McGown, Evelyn
AU - Henderson, L. M.
AU - Swan, Patricia B.
N1 - Funding Information:
This work was supported by Grant AM 13347-08 from the United States Public Health Service, Department of Health, Education, and Welfare and the Minnesota Agricultural Experiment Station. This paper is Scientific Journal Series No. 7696 , Minnesota Agricultural Experiment Station, University of Minnesota, St. Paul, Minn., 55101.
PY - 1971/12/30
Y1 - 1971/12/30
N2 - Interest in the use of the post-mitochondrial supernatant (PM-supernatant) for the study of in vitro amino acid incorporation arose from experiments indicating that the ribosomal aggregation of the PM-supernatant system more nearly represented that of the intact cell than did other ribosomal preparations. The PM-supernatant system was found to be 25 or 50 % more active than the microsomal and ribosomal amino acid incorporation systems. For PM-supernatant in vitro incorporation of l-[14C]valine into acid insoluble material, ATP, GTP, an energy generating system, Mg2+, K+, and a mixture of l-amino acids were necessary. Sephadex G-25 column chromatography of the PM-supernatant, pH 7.4, and presence of glutathione during homogenization of the tissue were necessary for optimal PM-supernatant in vitro incorporation. Under these conditions the rate of l-valine incorporation by the PM-supernatant from 0.17 to 0.20 nmoles/min per mg RNA for a liver from a fed rat. Fasting (20 h) prior to sacrifice, resulted in a 30-40 % decrease in amino acid incorporation by the PM-supernatant as well as a decrease in ribosomal aggregation. The decrease in PM-supernatant incorporation by fasting was found to be associated with both polysomal and cell sap factors.
AB - Interest in the use of the post-mitochondrial supernatant (PM-supernatant) for the study of in vitro amino acid incorporation arose from experiments indicating that the ribosomal aggregation of the PM-supernatant system more nearly represented that of the intact cell than did other ribosomal preparations. The PM-supernatant system was found to be 25 or 50 % more active than the microsomal and ribosomal amino acid incorporation systems. For PM-supernatant in vitro incorporation of l-[14C]valine into acid insoluble material, ATP, GTP, an energy generating system, Mg2+, K+, and a mixture of l-amino acids were necessary. Sephadex G-25 column chromatography of the PM-supernatant, pH 7.4, and presence of glutathione during homogenization of the tissue were necessary for optimal PM-supernatant in vitro incorporation. Under these conditions the rate of l-valine incorporation by the PM-supernatant from 0.17 to 0.20 nmoles/min per mg RNA for a liver from a fed rat. Fasting (20 h) prior to sacrifice, resulted in a 30-40 % decrease in amino acid incorporation by the PM-supernatant as well as a decrease in ribosomal aggregation. The decrease in PM-supernatant incorporation by fasting was found to be associated with both polysomal and cell sap factors.
UR - http://www.scopus.com/inward/record.url?scp=0015246547&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0015246547&partnerID=8YFLogxK
U2 - 10.1016/0005-2787(71)90881-1
DO - 10.1016/0005-2787(71)90881-1
M3 - Article
C2 - 5137607
AN - SCOPUS:0015246547
SN - 0005-2787
VL - 254
SP - 468
EP - 477
JO - BBA Section Nucleic Acids And Protein Synthesis
JF - BBA Section Nucleic Acids And Protein Synthesis
IS - 3
ER -