Hen oviduct N alpha-acetyltransferase is a ribonucleoprotein having 7 S RNA.
Hen oviduct N alpha-acetyltransferase was clarified to have a nucleic acid as an existing constituent by the following three results: (i) an ultraviolet absorption spectrum of the purified N alpha-acetyltransferase free of S-acetyl coenzyme A (Ac-CoA) had an absorption maximum at 260 nm. (ii) A nucleic acid band stained with ethidium bromide was detected on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. (iii) An ethidium bromide band co-migrated with a fluorescent band of the protein treated with N-(7-dimethylamino-4-methylcoumarinyl)maleimide, a reagent specific for thiol groups, on polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulfate. N alpha-Acetyltransferase lost its activity partially or completely by digestion with bovine pancreatic RNase A, Staphylococcus aureus nuclease, or proteinase K, showing that both the nucleic acid and the protein subunit were necessary for the enzyme activity. The nucleic acid component was identified as an RNA but not a DNA because the RNase T2 digest of the nucleic acid was composed of four 3'-ribomononucleotides and completely separated from 3'- and 5'-deoxyribomononucleotides on TLC. The chain length of the nucleic acid of 260 nucleotides estimated by formamide-polyacrylamide gel electrophoresis was calculated to be about 83,000 of the molecular weight. The contents of RNA (35.0%) and protein (65.0%) in N alpha-acetyltransferase determined on weight basis corresponded reasonably well to the contents of RNA (34.4%) and protein (65.6%) calculated based on the assumption that N alpha-acetyltransferase consisted of one molecule of 7 S RNA (Mr 83,000) and two identical Mr 79,000 protein subunits. The total molecular weight (241,000) of the holoenzyme calculated based on the above result was identical to the molecular weight (240,000) of N alpha-acetyltransferase estimated by Sepharose 6B gel filtration.
Kamitani
K
K
Division of Protein Chemistry, Osaka University, Japan.
Sakiyama
F
F
D000123:Acetyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002645:Chickens; D019286:Endopeptidase K; D005260:Female; D008836:Micrococcal Nuclease; D008970:Molecular Weight; D010057:Oviducts / Q000201:enzymology*; D012335:RNA, Ribosomal / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic; D012261:Ribonucleoproteins / Q000302:isolation & purification*; D012697:Serine Endopeptidases
D012335:RNA, Ribosomal; D012261:Ribonucleoproteins; D000123:Acetyltransferases; D012259:Ribonuclease, Pancreatic; D008836:Micrococcal Nuclease; D012697:Serine Endopeptidases; D019286:Endopeptidase K