Protein gene product 9.5 (PGP9.5) is a cytosolic protein that is highly expressed in vertebrate neurons, which is now included in the ubiquitin C-terminal hydrolase subclass (UCH) on the basis of primary-structure homology and hydrolytic activity on the synthetic substrate ubiquitin ethyl ester (UbOEt). Some UCHs show affinity for immobilized ubiquitin, a property exploited to purify them. In this study we show that this property can also be applied to PGP9.5, since a protein has been purified to homogeneity from bovine retina by affinity chromatography on a ubiquitin–Sepharose column that can be identified with: (a) PGP9.5 with respect to molecular mass, primary structure and immunological reactivity; (b) the known UCHs with respect to some catalytic properties, such as hydrolytic activity on UbOEt, (which also characterizes PGP9.5), Km value and reactivity with cysteine and histidine-specific reagents. However, it differs with respect to other properties, e.g. inhibition by UbOEt and a wider pH range of activity.
Affinity purification and characterization of protein gene product 9.5 (PGP9.5) from retina
Marco PICCININI, Adalberto MERIGHI, Renato BRUNO, Paolo CASCIO, Magda CURTO, Silvia Clarissa MIOLETTI, Maria T. RINAUDO; Affinity purification and characterization of protein gene product 9.5 (PGP9.5) from retina. Biochem J 1 September 1996; 318 (2): 711–716. doi: https://doi.org/10.1042/bj3180711
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