NTA center dot Ni2+-Functionalized Quantum Dots for VAMP2 Labeling in Live Cells

An efficient method for labeling individual proteins in live cells is required for investigations into biological mechanisms and cellular processes. Here we describe the preparation of small quantum dots (QDs) that target membrane surface proteins bearing a hexahistidine-tag (His(6)-tag) via specific binding to an nitrilotriacetic acid complex of nickel(II) (NTA center dot Ni2+) on the QD surfaces. We showed that the NTA center dot Ni2+-QDs bound to His-tag functionalized beads as a cellular mimic with high specificity and that QDs successfully targeted His(6)-tagged vesicle-associated membrane proteins (VMAP) on cell surfaces. This strategy provides an efficient approach to monitoring synaptic protein dynamics in spatially restricted and confined biological environments.This work was supported by grants from the Korea Science and Engineering Foundation (R01-2006-000- 10818-0) to S.C. funded by the Ministry of Education, Science and Technology, Republic of Korea.Dif A, 2009, J AM CHEM SOC, V131, P14738, DOI 10.1021/ja902743uUCHINOMIYA SH, 2009, CHEM COMMUN, V39, P5880Howarth M, 2008, NAT METHODS, V5, P397, DOI 10.1038/NMETH.1206Kim J, 2008, CHEM COMMUN, P1910, DOI 10.1039/b719434jLiu W, 2008, J AM CHEM SOC, V130, P1274, DOI 10.1021/ja076069pGroc L, 2007, J NEUROSCI, V27, P12433, DOI 10.1523/JNEUROSCI.3349-07.2007Jamieson T, 2007, BIOMATERIALS, V28, P4717, DOI 10.1016/j.biomaterials.2007.07.014MARIA ME, 2007, FEBS LETT, V581, P2905Lee IS, 2006, J AM CHEM SOC, V128, P10658, DOI 10.1021/ja063177nCourty S, 2006, NANO LETT, V6, P1491, DOI 10.1021/nl060921tMedintz IL, 2006, NAT MATER, V5, P581, DOI 10.1038/nmat1676Lata S, 2006, J AM CHEM SOC, V128, P2365, DOI 10.1021/ja0563105Howarth M, 2005, P NATL ACAD SCI USA, V102, P7583, DOI 10.1073/pnas.0503125102George N, 2004, J AM CHEM SOC, V126, P8896, DOI 10.1021/ja048396sGuignet EG, 2004, NAT BIOTECHNOL, V22, P440, DOI 10.1038/nbt954Xu CJ, 2004, J AM CHEM SOC, V126, P3392, DOI 10.1021/ja031776dLee KB, 2004, ANGEW CHEM INT EDIT, V43, P3048, DOI 10.1002/anie.200454088Dahan M, 2003, SCIENCE, V302, P442, DOI 10.1126/science.1088525Keppler A, 2003, NAT BIOTECHNOL, V21, P86, DOI 10.1038/nbt765Kapanidis AN, 2001, J AM CHEM SOC, V123, P12123, DOI 10.1021/ja017074aPathak S, 2001, J AM CHEM SOC, V123, P4103, DOI 10.1021/ja0058334Chen YA, 2001, NAT REV MOL CELL BIO, V2, P98, DOI 10.1038/35052017Hainfeld JF, 1999, J STRUCT BIOL, V127, P185, DOI 10.1006/jsbi.1999.4149KELLER TA, 1995, SUPRAMOL SCI, V2, P155, DOI 10.1016/0968-5677(96)89670-2GREEN NM, 1990, METHOD ENZYMOL, V184, P51PORATH J, 1975, NATURE, V258, P598, DOI 10.1038/258598a0