High-efficacy subcellular micropatterning of proteins using fibrinogen anchors.


Type
Article
Change log
Authors
Watson, Joseph L 
Aich, Samya 
Oller-Salvia, Benjamí 
Drabek, Andrew A 
Blacklow, Stephen C 
Abstract

Protein micropatterning allows proteins to be precisely deposited onto a substrate of choice and is now routinely used in cell biology and in vitro reconstitution. However, drawbacks of current technology are that micropatterning efficiency can be variable between proteins and that proteins may lose activity on the micropatterns. Here, we describe a general method to enable micropatterning of virtually any protein at high specificity and homogeneity while maintaining its activity. Our method is based on an anchor that micropatterns well, fibrinogen, which we functionalized to bind to common purification tags. This enhances micropatterning on various substrates, facilitates multiplexed micropatterning, and dramatically improves the on-pattern activity of fragile proteins like molecular motors. Furthermore, it enhances the micropatterning of hard-to-micropattern cells. Last, this method enables subcellular micropatterning, whereby complex micropatterns simultaneously control cell shape and the distribution of transmembrane receptors within that cell. Altogether, these results open new avenues for cell biology.

Description
Keywords
Animals, Cell Line, Fibrinogen, Green Fluorescent Proteins, Humans, Ligands, Microtechnology, Polyethylene Glycols, Subcellular Fractions
Journal Title
J Cell Biol
Conference Name
Journal ISSN
0021-9525
1540-8140
Volume Title
220
Publisher
Rockefeller University Press