New proteins involved in isoprenoid biosynthesis, useful in screening for inhibitors, also new intermediates, potential therapeutic agents, nucleic acids and antibodies

摘要

A protein (P1) capable of enzymatic conversion of 2C-methyl-D-erythritol-2,4-cyclodiphosphate (I) to 1-hydroxy-2-methyl-2-butenyl-4-diphosphate (II), especially the (E) form, is new. Independent claims are also included for: (1) a protein (P2) capable of enzymatic conversion of (II), especially the (E) form, to isopentenyldiphosphate (III) and/or dimethylallyl diphosphate (IV); (2) purified, isolated nucleic acid (NA) that encodes P1 or P2, with or without introns; (3) DNA expression vector that contains NA; (4) bacterial, protozoal, fungal, plant, insect or mammalian cells, cell cultures, organisms or their parts that are recombinantly provided with NA or the vector of (3); (5) cells, cell cultures, organisms or their parts transformed or transfected for increased formation of (II) or for increased conversion of (E)-(II) to (III) and/or (IV); (6) compounds, optionally isotopically labeled, of formula (X) and their salts; (7) pharmaceutical composition containing (X) and a carrier; (8) mono- or poly-clonal antibodies raised against (X); (9) method for detecting pathogens, preferably in body fluids, by using Ab; (10) method for altering expression levels of products of the ispG and/or ispH genes in cells; (11) method for identifying inhibitors of an enzyme that can convert (I) to(II) or (II) to (III) and (IV); (12) method for efficient in vivo synthesis of (II)-(IV), optionally isotopically labeled, in salt or protonated forms; (13) method of screening a chemical library for inhibitors of isoprenoid biosynthesis; (14) cells, cell cultures, organisms or their parts for efficient formation of biosynthetic products and intermediates of the 1-deoxy-D-xylulose-5-phosphate-dependent pathway; (15) method for efficient in vivo synthesis of 2C-methyl-D-erythritol-4-phosphate, 4-diphosphocytidyl-2C-methyl-D-erythritol (or its 2-phosphate), or (I)-(IV); (16) vector containing a recombinant sequence for use in cells of (14) or method (15); and (17) methods for chemical synthesis of (X). one of R1 and R2 = hydrogen, the other is -CH2O-P(O)(OH)-O-P(O)P(OH)2 or -CH2-O-P(O)(OH)2; and A = hydroxymethyl or formyl.