Rolling adhesion kinematics of yeast engineered to express selectins.
Selectins are cell adhesion molecules that mediate capture of leukocytes on vascular endothelium as an essential component of the inflammatory response. Here we describe a method for yeast surface display of selectins, together with a functional assay that measures rolling adhesion of selectin-expressing yeast on a ligand-coated surface. E-selectin-expressing yeast roll specifically on surfaces bearing sialyl-Lewis-x ligands. Observation of yeast rolling dynamics at various stages of their life cycle indicates that the kinematics of yeast motion depends on the ratio of the bud radius to the parent radius (B/P). Large-budded yeast "walk" across the surface, alternately pivoting about bud and parent. Small-budded yeast "wobble" across the surface, with bud pivoting about parent. Tracking the bud location of budding yeast allows measurement of the angular velocity of the yeast particle. Comparison of translational and angular velocities of budding yeast demonstrates that selectin-expressing cells are rolling rather than slipping across ligand-coated surfaces.
Bhatia
Sujata K
SK
Department of Bioengineering and Chemical Engineering, and Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Swers
Jeffrey S
JS
Camphausen
Raymond T
RT
Wittrup
K Dane
KD
Hammer
Daniel A
DA
D001696:Biomechanical Phenomena / Q000379:methods; D002448:Cell Adhesion / Q000502:physiology*; D020099:Coated Materials, Biocompatible / Q000378:metabolism; D019040:E-Selectin / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D009844:Oligosaccharides / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D012399:Rotation; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D000080506:Sialyl Lewis X Antigen
D020099:Coated Materials, Biocompatible; D019040:E-Selectin; D009844:Oligosaccharides; D011994:Recombinant Proteins; D000080506:Sialyl Lewis X Antigen