Identification of the three non‐identical subunits constituting human deoxyribonuclease II

FEBS Letters

Nakashima, Yoshimitsu¹  Mori, Shinjiro¹  Nakajima, Tamiko¹  Takeshita, Haruo¹  Kishi, Koichiro¹  Nomoto, Hiroshi³  Yasuda, Toshihiro¹  Hosomi, Osamu¹  Iida, Reiko² 

[1] Department of Legal Medicine, Gunma University School of Medicine, Maebashi 371-8511, Japan;Department of Legal Medicine, Fukui Medical School, Matsuoka-cho 910-1159, Japan;Laboratory of Molecular Biology, Gifu Pharmaceutical University, Gifu 502-8585, Japan

关键词: Amino acid sequence;    Deoxyribonuclease II;    Proteolytic processing;    Purification;    Subunit structure;    Human liver;    DTT;    dithiothreitol;    M;    mature protein;    2-ME;    2-mercaptoethanol;    P;    proprotein;    PT;    propeptide;    RT;    reverse transcriptase;    SRED;    single radial enzyme diffusion;    S;    signal peptide;   

DOI  :  10.1016/S0014-5793(98)01456-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown