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AAV serotypes have distinctive interactions with domains of the cellular receptor AAVR.

J. Virol.. 2017; 
PillaySirika,ZouWei,ChengFang,PuschnikAndreas S,MeyerNancy L,GanaieSafder S,DengXuefeng,WosenJonathan E,DavulcuOmar,YanZiying,EngelhardtJohn F,BrownKevin E,ChapmanMichael S,QiuJianming,CaretteJ
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The following antibodies were used in this study: mouse polyclonal anti-KIAA0319L(ab105385) from Abcam (Cambridge, CA), mouse monoclonal anti-FLAG antibody (F3165) from Sigma-Aldrich (St. Louis, MO), mouse monoclonal anti-GAPDH antibody (GT239) Genetex (Irvine, CA), mouse monoclonal anti-GST antibody (A00865) from Genscript(Piscataway, NJ), anti-intact AAV2 (clone A20, 03-61055) and anti-intact AAV5 antibody(clone ADK5a, 03-651148) from American Research Products, Inc. (Waltham, MA), antiLDLR (A304-417A) from Bethyl Laboratories, Inc. (Montgomery, TX), anti-ApoE2 (Cat# NB100-2216) from Novus Biologicals (Littleton, CO), anti-ORP150 (AF5558) from R&DSystems (Minneapolis, MN), and anti-integrin α5 antibody (AB1921) from EMD Millipore (Billerica, MA). Antibodies were used at dilutions as manufacturers suggested.

摘要

Adeno-associated virus (AAV) entry is determined by its interactions with specific surface glycans and proteinaceous receptor(s). Adeno-associated virus receptor (AAVR; also named KIAA0319L) is an essential cellular receptor required for the transduction of vectors derived from multiple AAV serotypes including the evolutionary distant serotypes, AAV2 and AAV5. Here, we further biochemically characterize the AAV-AAVR interaction and define the domains within the ectodomain of AAVR that facilitate this interaction. Using a virus overlay assay, it was previously shown that the major AAV2 binding protein in membrane preparations of human cells corresponds to a glycoprotein with a 150-kDa molecular mass. By es... More

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