Proteolytic Mapping of the Adeno-associated Virus Capsid

摘要: The three-dimensional structures of the viral capsid three AAV serotypes have previously been determined by X-ray crystallography or cryoelectron microscopy. These studies and similar autonomous parvoviruses yielded important structural information about virions in a low-energy conformation. However, there is little on properties solution under physiological conditions. We demonstrate that proteolytic digestion AAV2 with trypsin results cleavage at specific site surface while remains intact. products were mapped using unique antibodies, protein sequencing, mass spectroscopy, 3D structure modeling to region loop common all proteins. Empty capsids could be distinguished from full (DNA-containing) capsids, having an increased susceptibility VP2 being digested more rapidly chymotrypsin. Proteolytic analysis utilizing chymotrypsin was also capable distinguishing AAV1 AAV5, as seen differential fragment patterns. data novel approach for studying should valuable testing engineering vectors gene transfer.