De-mannosylation of phosphorylated N-glycans
First Claim
1. A method for demannosylating phosphorylated N-glycans on a glycoprotein, the method comprising contacting a glycoprotein comprising a terminal alpha-1,2 mannose linkage in which the underlying mannose is phosphorylated with a mannosidase, or a biologically active fragment of the mannosidase, that hydrolyzes a terminal alpha-1,2 mannose linkage when the underlying mannose is phosphorylated, wherein the contacting results in hydrolysis of the terminal alpha-1,2 mannose linkage, to produce a demannosylated glycoprotein, wherein the mannosidase is of the GH47 family or of the GH92 family.
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Abstract
Methods for demannosylating phosphorylated N-glycans on a glycoprotein are described that use a mannosidase capable of hydrolyzing a terminal alpha-1,2 mannose linkage when the underlying mannose is phosphorylated.
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Citations
37 Claims
- 1. A method for demannosylating phosphorylated N-glycans on a glycoprotein, the method comprising contacting a glycoprotein comprising a terminal alpha-1,2 mannose linkage in which the underlying mannose is phosphorylated with a mannosidase, or a biologically active fragment of the mannosidase, that hydrolyzes a terminal alpha-1,2 mannose linkage when the underlying mannose is phosphorylated, wherein the contacting results in hydrolysis of the terminal alpha-1,2 mannose linkage, to produce a demannosylated glycoprotein, wherein the mannosidase is of the GH47 family or of the GH92 family.
- 2. A method of producing a glycoprotein comprising demannosylated phosphorylated N-glycans, the method comprising introducing into a fungal cell a nucleic acid encoding the glycoprotein and the glycoprotein comprises a terminal alpha-1,2 mannose linkage in which the underlying mannose is phosphorylated, wherein the fungal cell is genetically engineered to comprise a nucleic acid encoding a mannosidase, or a biologically active fragment of the mannosidase, that hydrolyzes a terminal alpha-1,2 mannose linkage when the underlying mannose is phosphorylated, to produce a demannosylated glycoprotein, wherein the mannosidase is of the GH47 family or of the GH92 family.
Specification