CDNA CLONING OF A HUMAN 100 KDA DE-UBIQUITINATING ENZYME - THE 100 KDA HUMAN DE-UBIQUITINASE BELONGS TO THE UBIQUITIN C-TERMINAL HYDROLASE FAMILY-2 (UCH2)

The full length cDNA encoding a 100 kDa human de-ubiquitinating enzyme, referred to as de-ubiquitinase was obtained using one clone selected from a randomly sequenced human brain cDNA library and specific primers. The sequence of 18 peptides generated from the de-ubiquitinase isolated from outdated human erythrocytes matched perfectly with the predicted amino acid sequence, which would encode a protein containing 858 amino acids (calculated M(r) = 95,743 Da). Homology search disclosed that the protein is a member of a large family of ubiquitin C-terminal hydrolases (UCH2), that was defined on the basis of the presence of two specific patterns, 'the Cys- and His-domains', which are likely to be involved in the de-ubiquitinating activity [7]. An additional conserved region,'the aspartic acid domain', was also identified, the functional role of which is unknown.