The central interactive region of human MxA GTPase is involved in GTPase activation and interaction with viral target structures.
To define domains of the human MxA GTPase involved in GTP hydrolysis and antiviral activity, we used two monoclonal antibodies (mAb) directed against different regions of the molecule. mAb 2C12 recognizes an epitope in the central interactive region of MxA, whereas mAb M143 is directed against the N-terminal G domain. mAb 2C12 greatly stimulated MxA GTPase activity, suggesting that antibody-mediated crosslinking enhances GTP hydrolysis. In contrast, monovalent Fab fragments of 2C12 abolished GTPase activity, most likely by blocking intramolecular interactions required for GTPase activation. Interestingly, intact IgG molecules and Fab fragments of 2C12 both prevented association of MxA with viral nucleocapsids and neutralized MxA antiviral activity in vivo. mAb M143 had no effect on MxA function, indicating that this antibody binds outside functional regions. These data demonstrate that the central region recognized by 2C12 is critical for regulation of GTPase activity and viral target recognition.
Flohr
F
F
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008, Freiburg, Germany.
Schneider-Schaulies
S
S
Haller
O
O
Kochs
G
G
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000998:Antiviral Agents / Q000737:chemistry*; D001665:Binding Sites / Q000276:immunology; D015153:Blotting, Western; D002499:Centrifugation, Density Gradient; D034281:Dynamins; D004926:Escherichia coli / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000737:chemistry*; D019204:GTP-Binding Proteins; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D019251:Nucleocapsid / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding / Q000187:drug effects; D011506:Proteins / Q000737:chemistry* / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D011506:Proteins; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D034281:Dynamins