title: Single particle tracking of acyl carrier protein (ACP)-tagged TrkA receptors in PC12nnr5 cells.
abstract: There is a wide interest in studying the membrane mobility of Nerve Growth Factor (NGF) tropomyosin receptor kinase A (TrkA) at the single molecule level, in order to elucidate its diverse signaling responses related to different receptor functions. Here we present an experimental strategy based on the acyl carrier protein (ACP) tag in order to study the dynamics of the high-affinity NGF receptor TrkA in the membrane of PC12nnr5 cells. We present a single-particle tracking (SPT) study using highly photostable semiconductor quantum dots (Qdots) conjugated to ACP-tagged TrkA receptors. We demonstrate that ACP-TrkA shows biochemical and biological properties identical to those of its unmodified counterpart and that single receptor molecules in living cells display distinct diffusive regimes and a highly heterogeneous dynamics.
authors: NEST Laboratory, Scuola Normale Superiore and Istituto Nanoscienze - CNR, Piazza San Silvestro 12, I-56126 Pisa, Italy; IIT@NEST, Center for Nanotechnology Innovation, Piazza San Silvestro 12, I-56126 Pisa, Italy; Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy. Electronic address: a.callegari@sns.it.
authors: Andrea
authors: A
authors: Callegari
authors: NEST Laboratory, Scuola Normale Superiore and Istituto Nanoscienze - CNR, Piazza San Silvestro 12, I-56126 Pisa, Italy; Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy.
authors: Stefano
authors: S
authors: Luin
authors: NEST Laboratory, Scuola Normale Superiore and Istituto Nanoscienze - CNR, Piazza San Silvestro 12, I-56126 Pisa, Italy; Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy.
authors: Laura
authors: L
authors: Marchetti
authors: Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy.
authors: Alessandro
authors: A
authors: Duci
authors: Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy.
authors: Antonino
authors: A
authors: Cattaneo
authors: NEST Laboratory, Scuola Normale Superiore and Istituto Nanoscienze - CNR, Piazza San Silvestro 12, I-56126 Pisa, Italy; IIT@NEST, Center for Nanotechnology Innovation, Piazza San Silvestro 12, I-56126 Pisa, Italy; Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56127 Pisa, Italy.
authors: Fabio
authors: F
authors: Beltram
mesh_terms: D000213:Acyl Carrier Protein / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D015336:Molecular Probe Techniques; D016716:PC12 Cells; D011471:Prostatic Neoplasms / Q000378:metabolism*; D045663:Quantum Dots; D051381:Rats; D020917:Receptor, trkA / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D000213:Acyl Carrier Protein; D020917:Receptor, trkA