Surface binding kinetics of prothrombin fragment 1 on planar membranes measured by total internal reflection fluorescence microscopy

Biochemistry ◽  
1992 ◽  
Vol 31 (26) ◽  
pp. 5983-5995 ◽  
Author(s):  
Kenneth H. Pearce ◽  
Richard G. Hiskey ◽  
Nancy L. Thompson
Keyword(s):  
Fluorescence Microscopy ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Binding Kinetics ◽  
Surface Binding ◽  
Prothrombin Fragment ◽  
Planar Membranes ◽  
Kinetics Of

scholarly journals Imaging Constitutive Exocytosis with Total Internal Reflection Fluorescence Microscopy

2000 ◽  
Vol 149 (1) ◽  
pp. 23-32 ◽  
Author(s):  
Jan Schmoranzer ◽  
Mark Goulian ◽  
Dan Axelrod ◽  
Sanford M. Simon
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Fluorescence Microscopy ◽  
Total Internal Reflection ◽  
Fluorescent Protein ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Green Fluorescent ◽  
Kinetics Of ◽  
Flattening Process

Total internal reflection fluorescence microscopy has been applied to image the final stage of constitutive exocytosis, which is the fusion of single post-Golgi carriers with the plasma membrane. The use of a membrane protein tagged with green fluorescent protein allowed the kinetics of fusion to be followed with a time resolution of 30 frames/s. Quantitative analysis allowed carriers undergoing fusion to be easily distinguished from carriers moving perpendicularly to the plasma membrane. The flattening of the carriers into the plasma membrane is seen as a simultaneous rise in the total, peak, and width of the fluorescence intensity. The duration of this flattening process depends on the size of the carriers, distinguishing small spherical from large tubular carriers. The spread of the membrane protein into the plasma membrane upon fusion is diffusive. Mapping many fusion sites of a single cell reveals that there are no preferred sites for constitutive exocytosis in this system.

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