Transactivator of transcription fusion protein transduction causes membrane inversion.
The transactivator of transcription (TAT) protein transduction domain is an 11-amino acid positively charged peptide that has been shown to pull diverse molecules across cell membranes in vitro and in vivo. Fusion proteins constructed with TAT rapidly enter and exit cells and have been shown to cross intracellular membranes as well. Electrostatic interactions between TAT and the cell membrane have been implicated as a part of the mechanism of transduction. Here, we report that TAT transduction causes membrane phospholipid rearrangement as evidenced by detection of phosphatidylserine on the outer surface of the cell membrane. Furthermore, these rearrangements can be blocked by positively charged polylysine, further implicating electrostatic interactions as a part of the mechanism. Neither apoptosis nor necrosis is induced in these cells after exposure to TAT. We conclude that the process of TAT.GFP transduction causes phosphatidylserine to translocate from the inner to the outer leaflet of the plasma membrane. These results provide insight into the mechanism of TAT protein transduction domain transduction.
Del Gaizo Moore
Victoria
V
Division of Molecular Medicine, Department of Internal Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157-1081, USA.
Payne
R Mark
RM
D000818:Animals; D017302:Annexins / Q000737:chemistry; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D003094:Collagen / Q000378:metabolism; D004396:Coloring Agents / Q000737:chemistry; D004789:Enzyme Activation; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009336:Necrosis; D016716:PC12 Cells; D010718:Phosphatidylserines / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D055672:Static Electricity; D015533:Transcriptional Activation
D017302:Annexins; D004396:Coloring Agents; D008164:Luminescent Proteins; D010718:Phosphatidylserines; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011107:Polylysine; D003094:Collagen; D020169:Caspases