Molecular construction of bionanoparticles: chimaeric SIV p17-HIV I p6 nanoparticles with minimal viral protein content.
VLPs (virus-like particles) are promising delivery vectors for molecular therapy, since they combine the major advantages of viral vectors with significantly fewer viral vector disadvantages. The present paper describes the molecular construction of chimaeric VLPs based on minimal SIV (simian immunodeficiency virus) and HIV1 components. A chimaeric protein was constructed by fusion of SIV matrix protein (p17) and HIV1 p6 protein, and we demonstrated that the chimaeric proteins assemble as 80 nm nanoparticles containing approximately 7700 chimaeric protein units. Chimaeric VLPs are released from HEK-293T cells (human embryonic kidney cells expressing the large T-antigen of simian virus 40) and are fully encapsulated with lipid membrane. Chimaeric VLPs are produced at 3.7-fold higher levels when compared with SIV p17 VLPs owing to duplication of a PTAP (Pro-Thr-Ala-Pro) domain previously shown as essential for virus particle release. The chimaeric VLPs constructed in the present paper were efficiently pseudotyped with vesicular-stomatitis-virus glycoprotein, as shown by immunoprecipitation assays.
Costa
Maria J L
MJ
URIA (Unidade dos Retrovírus e Infecções Associadas), Faculdade de Farmácia Universidade de Lisboa, Lisbon, Portugal.
Pedro
Luísa
L
de Matos
António P Alves
AP
Aires-Barros
Maria R
MR
Belo
José A
JA
Goncalves
João
J
Ferreira
Guilherme N M
GN
D002460:Cell Line; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D015202:Protein Engineering / Q000379:methods*; D013539:Simian virus 40 / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry; D014771:Virion / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D014764:Viral Proteins