Monomeric Forms of Human Aminoacyl-tRNA Synthetases Having Non-Canonical Biological Activities

Abstract

Isolated monomeric aminoacyl-tRNA synthetase polypeptides and polynucleotides having non-canonical biological activities are provided, as well as compositions and methods related thereto.

What is claimed is: 
     
         1 . An isolated aminoacyl-tRNA (AARS) synthetase polypeptide having a non-canonical biological activity, or an active fragment or variant thereof, wherein the polypeptide is substantially in a monomeric form under physiological conditions or in solution. 
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the polypeptide comprises one or more stabilizing modifications in relation to a wild-type AARS sequence that reduce its ability to dimerize with itself or another AARS polypeptide. 
     
     
         3 . The isolated polypeptide of  claim 2 , wherein the one or more modifications are selected from amino acid substitution(s), amino acid deletion(s), amino acid addition(s)/insertion(s), truncation(s), and chemical modification(s). 
     
     
         4 . The isolated polypeptide of  claim 2 , comprising one or more stabilizing modifications located within or proximal to the primary, secondary, or tertiary structure of the dimer interface. 
     
     
         5 . The isolated polypeptide of  claim 2 , wherein the one or more stabilizing modifications are proximal to the primary structure of the dimer interface by about 1-20 residues. 
     
     
         6 . The isolated polypeptide of any one of  claims 1 - 5 , wherein the polypeptide is a substantially monomeric form of tyrosyl-tRNA synthetase (YRS), tryptophanyl-tRNA synthetase (WRS), lysyl-tRNA synthetase (KRS), glutaminyl-tRNA synthetase (QRS), glycyl-tRNA synthetase (GlyRS), histidyl-tRNA synthetase (HisRS), seryl-tRNA synthetase (SRS), phenyl alanyl-tRNA synthetase (PheRS), alanyl-tRNA synthetase (AlaRS), asparaginyl-tRNA synthetase (AsnRS), aspartyl-tRNA synthetase (AspRS), cysteinyl-tRNA synthetase (CysRS), glutamyl-tRNA synthetase (ERS), prolyl-tRNA synthetase (ProRS), arginyl-tRNA synthetase (RRS), isoleucyl-tRNA synthetase (IRS), leucyl-tRNA synthetase (LRS), threonyl-tRNA synthetase (TRS), methionyl-tRNA synthetases (MRS), or valyl-tRNA synthetase (VRS). 
     
     
         7 . The isolated polypeptide of  claim 6 , wherein the polypeptide is a tyrosyl-tRNA (YRS) synthetase polypeptide. 
     
     
         8 . The isolated polypeptide of  claim 7 , comprising one or more stabilizing modifications of Pro 159, Leu 160, or Leu 161, or any combination thereof. 
     
     
         9 - 12 . (canceled) 
     
     
         13 . The isolated polypeptide of  claim 6 , wherein the polypeptide is a tryptophanyl-tRNA synthetase (WRS) polypeptide. 
     
     
         14 . The isolated polypeptide of  claim 13 , comprising one or more stabilizing modifications of F260, Y201, I278, H130, or E408, or any combination thereof. 
     
     
         15 . The isolated polypeptide of  claim 13 , comprising an amino acid sequence at least 80%, 85%, 90%, 95%, or 98% identical to SEQ ID NO:4 or 7-9, or a biologically active fragment thereof, wherein the polypeptide comprises a modification of F260, Y201, I278, H130, or E408, or any combination thereof. 
     
     
         16 . The isolated polypeptide of  claim 13 , comprising the amino acid sequence of SEQ ID NO:8 or 9. 
     
     
         17 . The isolated polypeptide of  claim 6 , wherein the polypeptide is a lysyl-tRNA synthetase (KRS) polypeptide, or a truncation or variant thereof. 
     
     
         18 - 34 . (canceled) 
     
     
         35 . A pharmaceutical composition, comprising an isolated aminoacyl-tRNA (AARS) synthetase polypeptide according to any one of  claims 1 - 34 , and a pharmaceutically acceptable carrier. 
     
     
         36 . A method of modulating an inflammatory response in subject, comprising administering to the subject an isolated aminoacyl-tRNA (AARS) synthetase polypeptide according to any one of  claims 1 - 34 , or a pharmaceutical composition according to  claim 15 . 
     
     
         57 . A method of treating a condition, comprising administering to a subject in need thereof an isolated aminoacyl-tRNA (AARS) synthetase polypeptide according to any one of  claims 1 - 34 , or a pharmaceutical composition according to  claim 35 , wherein the condition is selected from the group consisting of an inflammatory condition, an autoimmune condition, a neoplastic disease, a metabolic disease, a neurological disease, an infection, an immunodeficiency, a cardiovascular disease, and a condition associated with angiogenesis. 
     
     
         65 . A composition, comprising a human immunodeficiency virus (HIV) Gag protein and a substantially monomeric lysyl tRNA synthetase (KRS) polypeptide according to any one of  claims 17 - 26 , which comprises one or more stabilizing modifications that do not significantly reduce KRS binding to HIV Gag. 
     
     
         66 . A modified cell, comprising a human immunodeficiency virus (HIV) Gag protein and an exogenous, substantially monomeric lysyl tRNA synthetase (KRS) polypeptide according to any one of  claims 17 - 26 , which comprises one or more stabilizing modifications that do not significantly reduce KRS binding to HIV Gag. 
     
     
         74 . A compound or binding agent that specifically binds to (a) an H3 and/or H4 region of a human immunodeficiency virus (HIV) Gag-CA-CTD domain, (b) an inter-domain cavity of substantially monomeric KRS which interacts with the H3 and/or H3 region of Gag-CA-CTD domain, or both (a) and (b), and reduces the interaction between HIV Gag and monomeric KRS. 
     
     
         75 - 76 . (canceled) 
     
     
         77 . A method of reducing assembly of a human immunodeficiency virus (HIV), comprising contacting an HIV-infected cell with a compound identified by the method of any one of claims  69 - 73 , or the compound or binding agent according to any one of  claims 74 - 76 . 
     
     
         78 - 105 . (canceled)