Assignment of A and B types of monoamine oxidase in NCB20 hybrid cells to those of the parental cells by peptide mapping.
The structures of [3H]pargyline-labeled, flavin-containing polypeptides of monoamine oxidase (MAO) from hybrid NCB20 cells, and their parental cells, A/J mouse brain cells and Chinese hamster brain cells, were analyzed and compared by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and limited proteolysis and one-dimensional peptide mapping in SDS gels. After preincubation of mitochondrial preparations with deprenyl or clorgyline, the flavin-containing polypeptide of type A or type B MAO was selectively labeled with [3H]pargyline. SDS-PAGE of [3H]pargyline-labeled mitochondrial samples revealed that the polypeptide with apparent Mr of 62,000 was associated with type A activity in the three types of cells, and that the polypeptide with apparent Mr of 61,000 or 58,000 was associated with type B activity in Chinese hamster brain cells and NCB20 cells or A/J mouse brain cells, respectively. Chymotrypsin digestion of the [3H]pargyline-labeled polypeptides and the peptide mapping in SDS gels from A/J mouse and Chinese hamster brain cells produced identical map patterns between the two type A MAOs, almost the same map patterns (with the exception of one additional peptide fragment) between the two type B MAOs, and different map patterns between type A and type B MAOs. The results of identical treatments of the [3H]pargyline-labeled polypeptides of MAOs in NCB20 cells showed that type A and type B MAO in NCB20 cells were similar to type A MAO of A/J mouse and Chinese hamster brain cells and to type B MAO of Chinese hamster brain cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Nakano
T
T
Saito
S
S
Higashida
H
H
Kojima
K
K
Nagatsu
T
T
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000201:enzymology*; D002460:Cell Line; D002918:Chymotrypsin / Q000378:metabolism; D003010:Clorgyline / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006822:Hybrid Cells / Q000201:enzymology*; D051379:Mice; D008805:Mice, Inbred A; D008928:Mitochondria / Q000201:enzymology; D008970:Molecular Weight; D008995:Monoamine Oxidase / Q000378:metabolism*; D010293:Pargyline / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D012642:Selegiline / Q000494:pharmacology
D010446:Peptide Fragments; D005182:Flavin-Adenine Dinucleotide; D012642:Selegiline; D010293:Pargyline; D008995:Monoamine Oxidase; D002918:Chymotrypsin; D003010:Clorgyline