Exploring Transduction Mechanisms of Protein Transduction Domains (PTDs) in Living Cells Utilizing Single-Quantum Dot Tracking (SQT) Technology.docVIP

Sensors 2012, 12, 549-572; doi:10.3390/s120100549 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Review Exploring Transduction Mechanisms of Protein Transduction Domains (PTDs) in Living Cells Utilizing Single-Quantum Dot Tracking (SQT) Technology Yasuhiro Suzuki Department of Emerging Infectious Diseases, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan; E-Mail: suzukiy39@med.tohoku.ac.jp; Tel: +81-022-717-8220; Fax: +81-022-717-8221 Received: 30 November 2011; in revised form: 30 December 2011 / Accepted: 31 December 2011 / Published: 5 January 2012 Abstract: Specific protein domains known as protein transduction domains (PTDs) can permeate cell membranes and deliver proteins or bioactive materials into living cells. Various approaches have been applied for improving their transduction efficacy. It is, therefore, crucial to clarify the entry mechanisms and to identify the rate-limiting steps. Because of technical limitations for imaging PTD behavior on cells with conventional fluorescent-dyes, how PTDs enter the cells has been a topic of much debate. Utilizing quantum dots (QDs), we recently tracked the behavior of PTD that was derived from HIV-1 Tat (TatP) in living cells at the single-molecule level with 7-nm special precision. In this review article, we initially summarize the controversy on TatP entry mechanisms; thereafter, we will focus on our recent findings on single-TatP-QD tracking (SQT), to identify the major sequential steps of intracellular delivery in living cells and to discuss how SQT can easily provide direct information on TatP entry mechanisms. As a primer for SQT study, we also discuss the latest findings on single particle tracking of various molecules on the plasma membrane. Finally, we discuss the problems of QDs and the challenges for the future in utilizing currently available