Enhancement of biological reactions on cell surfaces via macromolecular crowding

Rafi Chapanian, David H. Kwan, Iren Constantinescu, Fathima A. Shaikh, Nicholas A..A. Rossi, Stephen G Withers () and Jayachandran N Kizhakkedathu ()
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Rafi Chapanian: Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
David H. Kwan: University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1
Iren Constantinescu: Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
Fathima A. Shaikh: University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1
Nicholas A..A. Rossi: Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
Stephen G Withers: University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1
Jayachandran N Kizhakkedathu: Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3

Nature Communications, 2014, vol. 5, issue 1, 1-12

Abstract: Abstract The reaction of macromolecules such as enzymes and antibodies with cell surfaces is often an inefficient process, requiring large amounts of expensive reagent. Here we report a general method based on macromolecular crowding with a range of neutral polymers to enhance such reactions, using red blood cells (RBCs) as a model system. Rates of conversion of type A and B red blood cells to universal O type by removal of antigenic carbohydrates with selective glycosidases are increased up to 400-fold in the presence of crowders. Similar enhancements are seen for antibody binding. We further explore the factors underlying these enhancements using confocal microscopy and fluorescent recovery after bleaching (FRAP) techniques with various fluorescent protein fusion partners. Increased cell-surface concentration due to volume exclusion, along with two-dimensionally confined diffusion of enzymes close to the cell surface, appear to be the major contributing factors.

Date: 2014
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DOI: 10.1038/ncomms5683

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