TY - JOUR
T1 - Characterizing fluorescence recovery curves for nuclear proteins undergoing binding events
AU - Carrero, G.
AU - Crawford, E.
AU - Hendzel, M. J.
AU - De Vries, G.
N1 - Funding Information:
This research was supported in part by MITACS, a Canadian Network of Centers of Excellence (GC) and the Natural Sciences and Engineering Research Council of Canada (GC and GdeV). MJH is supported by scholarships from the Alberta Heritage Foundation for Medical Research and the Canadian Institutes of Health Research, and project funding was provided by the Alberta Cancer Board.
PY - 2004/11
Y1 - 2004/11
N2 - Fluorescence recovery after photobleaching (FRAP) is an experimental technique used to measure the mobility of proteins within the cell nucleus. After proteins of interest are fluorescently tagged for their visualization and monitoring, a small region of the nucleus is photobleached. The experimental FRAP data are obtained by recording the recovery of the fluorescence in this region over time. In this paper, we characterize the fluorescence recovery curves for diffusing nuclear proteins undergoing binding events with an approximate spatially homogeneous structure. We analyze two mathematical models for interpreting the experimental FRAP data, namely a reaction-diffusion model and a compartmental model. Perturbation analysis leads to a clear explanation of two important limiting dynamical types of behavior exhibited by experimental recovery curves, namely, (1) a reduced diffusive recovery, and (2) a biphasic recovery characterized by a fast phase and a slow phase. We show how the two models, describing the same type of dynamics using different approaches, relate and share common ground. The results can be used to interpret experimental FRAP data in terms of protein dynamics and to simplify the task of parameter estimation. Application of the results is demonstrated for nuclear actin and type H1 histone.
AB - Fluorescence recovery after photobleaching (FRAP) is an experimental technique used to measure the mobility of proteins within the cell nucleus. After proteins of interest are fluorescently tagged for their visualization and monitoring, a small region of the nucleus is photobleached. The experimental FRAP data are obtained by recording the recovery of the fluorescence in this region over time. In this paper, we characterize the fluorescence recovery curves for diffusing nuclear proteins undergoing binding events with an approximate spatially homogeneous structure. We analyze two mathematical models for interpreting the experimental FRAP data, namely a reaction-diffusion model and a compartmental model. Perturbation analysis leads to a clear explanation of two important limiting dynamical types of behavior exhibited by experimental recovery curves, namely, (1) a reduced diffusive recovery, and (2) a biphasic recovery characterized by a fast phase and a slow phase. We show how the two models, describing the same type of dynamics using different approaches, relate and share common ground. The results can be used to interpret experimental FRAP data in terms of protein dynamics and to simplify the task of parameter estimation. Application of the results is demonstrated for nuclear actin and type H1 histone.
UR - http://www.scopus.com/inward/record.url?scp=7444240826&partnerID=8YFLogxK
U2 - 10.1016/j.bulm.2004.02.005
DO - 10.1016/j.bulm.2004.02.005
M3 - Journal Article
C2 - 15522344
AN - SCOPUS:7444240826
SN - 0092-8240
VL - 66
SP - 1515
EP - 1545
JO - Bulletin of Mathematical Biology
JF - Bulletin of Mathematical Biology
IS - 6
ER -