Methods for ultrafast formation of disulfide bonds in peptides and proteins
AssigneeTechnion Research & Development Foundation Limited
InventorsAshraf BRIK, Shay LAPS, Fatima ATAMLEH, Guy KAMNESKY
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Abstract
Provided herein a method for forming one or more intramolecular disulfide bonds.
Claims40
A method for forming a plurality of disulfide bonds, comprising
- a. providing a peptide comprising (i) a deprotected cysteine pair, (ii) a first cysteine pair protected by a first protecting group, and optionally (iii) a second cysteine pair protected by a second protecting group;
- b. adding to said peptide a sufficient amount of a reagent comprising a disulfide, a diselenide, a thiol, a sulfide, a thiocarbonate, a dithiocarbonate, thiocarbamate, or a dithiocarbamate, including any combination, a disulfanyl derivate, a metal complex or a salt thereof, thereby forming a first disulfide bond between thiols of said deprotected cysteine pair;
- c. providing said peptide under conditions sufficient for deprotecting the first protecting group and adding a sufficient amount of said reagent to said peptide, thereby forming a second disulfide bond between thiols of said first cysteine pair;
- wherein said plurality of disulfide bonds comprises intramolecular bonds;
- and wherein the steps b and c are performed in one pot.
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A method for forming a disulfide bond, comprising:
- a. providing a peptide comprising a first cysteine pair, wherein each thiol of said first cysteine pair is protected by a photocleavable protecting group;
- b. exposing said peptide to light for a time period sufficient for substantially deprotecting said photocleavable protecting group;
- c. adding a sufficient amount of a reagent comprising a disulfide, a diselenide, a thiol, a sulfide, a thiocarbonate, a dithiocarbonate, thiocarbamate, or a dithiocarbamate, including any combination, a disulfanyl derivate, a metal complex or a salt thereof, thereby forming a first disulfide bond between thiols of the first cysteine pair;
- wherein said disulfide bond is an intramolecular bond;
- and wherein the step b and the step c are performed simultaneously or subsequently.
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A composition comprising a peptide comprising one or more intramolecular disulfide bond, and a residual amount of a reagent comprising a disulfide, a diselenide, a thiol, a sulfide, a thiocarbonate, a dithiocarbonate, thiocarbamate, or a dithiocarbamate, including any combination, a disulfanyl derivate, a metal complex or a salt thereof.
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Dependencies are inferred from the claim text.