Membrane-associated human tyrosinase is an enzymatically active monomeric glycoprotein.

Membrane-associated human tyrosinase is an enzymatically active monomeric glycoprotein.

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Human tyrosinase (hTyr) is a Type 1 membrane bound glycoenzyme that catalyzes the initial and rate-limiting steps of melanin production in the melanosome.Mutations in the Tyr gene are linked to oculocutaneous albinism type 1 (OCA1), an autosomal recessive disorder.Currently, the application of enzyme replacement therapy for a treatment of OCA1 is hampered Pour Test Kits by the absence of pure hTyr.Here, full-length hTyr (residues 1-529) was overexpressed in Trichoplusia ni larvae infected with a baculovirus, solubilized with detergent and purified using chromatography.Michaelis-Menten kinetics, enzymatic specific activity, and analytical ultracentrifugation were used to compare the hTyr in detergent with the soluble recombinant intra-melanosomal domain, hTyrCtr (residues 19-469).

Active hTyr is monomeric in detergent micelles suggesting no stable interactions between protein molecules.Both, hTyr and Baby Bodysuit hTyrCtr, exhibited similar enzymatic activity and ligand affinity in L-DOPA and L-Tyrosine reactions.In addition, expression in larvae is a scalable process that will allow high yield protein production.Thus, larval production of enzymatically active human tyrosinase potentially could be a useful tool in developing a cure for OCA1.