On the possible involvement of bovine serum albumin precursor in lipofection pathway.
Protein factors involved in lipofection pathways remain elusive. Using avidin-biotin affinity chromatography and mass finger printing analysis technique, herein we report the identification of a 70 kDa size protein (bovine serum albumin precursor, BSAP) which binds strongly with lipoplexes and may play role in lipofection pathway. Using multiple cultured animal cells and three structurally different cationic transfection lipids, we show that the efficiencies of liposomal transfection vectors get significantly enhanced (by ~2.5- to 5.0-fold) in cells pre-transfected with lipoplexes of reporter plasmid construct encoding BSAP. Findings in the cellular uptake experiments in A549 cells cultured in DMEM supplemented with 10 percent (w/w) BODIPY-labelled BSAP are consistent with the supposition that BSAP enters cell cytoplasm from the cell culture medium (DMEM supplemented with 10 percent FBS) used in lipofection. Cellular uptake studies by confocal microscopy using BODIPY-labelled BSAP and FITC-labelled plasmid DNA revealed co-localization of plasmid DNA and BSAP within the cell cytoplasm and nucleus. In summary, the present findings hint at the possible involvement of BSAP in lipofection pathway.
Biomaterials Group, CSIR-Indian Institute of Chemical Technology, Hyderabad 500 007, India.
Anubhab
A
Mukherjee
Jayanta
J
Bhattacharyya
Arabinda
A
Chaudhuri
D000818:Animals; D001360:Avidin; D001710:Biotin; D001896:Boron Compounds; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D016172:DNA Fingerprinting; D005293:Ferritins; D006801:Humans; D008081:Liposomes / Q000378:metabolism*; D018613:Microscopy, Confocal; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D008081:Liposomes; D011498:Protein Precursors; C051805:avidin-ferritin biotin; D001360:Avidin; D012710:Serum Albumin, Bovine; D001710:Biotin; D005293:Ferritins