Identification of the three non-identical subunits constituting human deoxyribonuclease II.
We purified DNase II from human liver to apparent homogeneity. The N-terminal amino acid sequences of each of three components constituting the purified mature enzyme were then separately determined by automatic Edman degradation. A combination of this chemical information and the previously reported nucleotide sequence of the cDNA encoding human DNase II [Yasuda et al. (1998) J. Biol. Chem. 273, 2610-2626] allowed detailed elucidation of the enzyme's subunit structure: human DNase II was composed of three non-identical subunits, a propeptide, proprotein and mature protein, following a signal peptide. Expression analysis of a series of deletion mutants derived from the cDNA of DNase II in COS-7 cells suggested that although a single large precursor protein may not be necessary for proteolytic maturation, the propeptide region L17-Q46 may play an essential role in generating the active form of the enzyme.
Takeshita
H
H
Department of Legal Medicine, Gunma University School of Medicine, Maebashi, Japan.
Yasuda
T
T
Iida
R
R
Nakajima
T
T
Hosomi
O
O
Nakashima
Y
Y
Mori
S
S
Nomoto
H
H
Kishi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D008099:Liver / Q000201:enzymology; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation; D021382:Protein Sorting Signals; D017421:Sequence Analysis; D017384:Sequence Deletion; D014162:Transfection
D004792:Enzyme Precursors; D021382:Protein Sorting Signals; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II