Patent ID: 6376253
Filing Date: 2002-04-23
Classification: C07B,C07K,C12N,C12P,G01R,Y10T

Abstract:
A method for determining three-dimensional structure information of a peptidic sequence of 50 or more amino acids, comprising:(a) providing an isotopically substituted nutrient medium which contains all amino acids required for protein biosynthesis, assimilable sources of carbohydrate, essential minerals and growth factors, wherein at least one species of said amino acids in said nutrient medium contains an isotopic substitution; wherein said isotopic substitution is in the backbone structure but not the side chains of said at least one species of amino acid or in the hydrogen atoms bonded to the &agr;-carbon of said at least one species of amino acid or both; and wherein said isotopic substitution is selected from 13C; 15N; 2H; 13C and 2H; 15N and 2H; 13C and 15N; and 13C, 15N and 2H; with the proviso that when said at least one species of amino acid is glycine, isotopic substitutions of 2H may occur at either or both of the hydrogen atoms bonded to the &agr;-carbon with the proviso that essentially none of the isotopic substitutions are present on the side chains of any amino acid; (b) cultivating in said isotopically substituted nutrient medium, under protein-producing conditions, a bacterial, yeast, mammalian or insect cell culture capable of producing said peptidic sequence of 50 or more amino acids; (c) recovering said isotopically substituted peptide sequence of 50 or more amino acids from said nutrient medium; and (d) subjecting said isotopically substituted peptidic sequence of 50 or more amino acids to NMR spectroscopic analysis to determine information about its three-dimensional structure.