Proteins, methods, and systems for subcellular localization of proteins for post-translation modification
AssigneeBetter Dairy Limited
InventorsBernd Alfons Gerhartz, Kate Royle, Christian Arenas López, Camila Aparecida Cotrim, Agata Giardina
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Abstract
The present disclosure provides for proteins, methods, systems for directing enzymes for generating post-translational modifications on proteins to a particular subcellular location. Specifically, kinases for phosphorylation of proteins containing one or more sites susceptible to phosphorylation are disclosed, also with methods and systems for phosphorylating proteins of interest, such as proteins having nutritional and therapeutic uses.
Claims21
-107. (canceled)
A method comprising:
- (a) providing a recombinant host cell comprising:
- i) a non-naturally occurring polypeptide comprising a serine/threonine kinase coupled to a heterologous anchoring domain, wherein the anchoring domain is capable of anchoring the serine/threonine kinase to an intracellular membrane of the recombinant host cell;
- and
- ii) a protein heterologous to the recombinant host cell;
- and
- (b) using the non-naturally occurring polypeptide to phosphorylate the protein heterologous to the recombinant host cell, thereby generating a phosphorylated protein.
A non-naturally occurring polypeptide comprising a serine/threonine kinase coupled to a heterologous domain capable of anchoring the serine/threonine kinase to an intracellular membrane of a cell.
A composition comprising phosphorylated non-naturally occurring secretory polypeptides, wherein the phosphorylated non-naturally occurring secretory polypeptides comprise a higher level of phosphorylation than corresponding non-naturally occurring secretory polypeptides phosphorylated in a recombinant host cell using Fam20c kinase without a heterologous anchoring domain, wherein the phosphorylated non-naturally occurring secretory polypeptides comprise a osteopontin polypeptide or a casein polypeptide.
Dependencies are inferred from the claim text.