Cloning, expression and characterisation of murine procathepsin E

FEBS Letters

Tatnell, Peter J¹  Lees, Wendy E¹  Kay, John¹ 

[1] School of Molecular and Medical Biosciences, University of Wales, Cardiff, P.O. Box 911, Cardiff, CF1 3US Wales, UK

关键词: Murine procathepsin E;    cloning;    Murine procathepsin E;    expression in E. coli;    Recombinant cathepsin E;    Characterization;    Chromogenic substrate hydrolysis;    inhibition;    RT-PCR;    reverse transcriptase polymerase chain reaction;    RACE;    rapid amplification of cDNA ends;   

DOI  :  10.1016/S0014-5793(97)00388-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNA encoding murine procathepsin E was isolated and sequenced and recombinant enzyme was produced in Escherichia coli. The activity of the purified recombinant mouse cathepsin E was characterised quantitatively using two synthetic peptide substrates and naturally occurring inhibitors. The majority of the recombinant enzyme was present as a homodimer (M_r ∼80) in which the two monomers were linked by an intermolecular disulfide bond. By analogy to previous studies with human cathepsin E, this is most likely a consequence of the presence of a unique cysteine residue near the N-terminus of the mature proteinase. The availability of (i) recombinant murine enzyme in reasonable quantities and (ii) a full-length cDNA now enables structural investigations and attempts to generate ‘knock-out’ mice deficient in this important aspartic proteinase to be undertaken.

【 授权许可】

Unknown