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Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells.

Publication ,  Journal Article
Chan, BP; Chilkoti, A; Reichert, WM; Truskey, GA
Published in: Biomaterials
February 2003

We have previously shown that the high-affinity streptavidin (SA)-biotin interaction enhanced the initial integrin-mediated adhesion of biotinylated endothelial cells to SA-coated surface by serving as an extrinsic bond to stabilize and enhance the intrinsic fibronectin-integrin binding between the cell and surface. However, the SA-biotin interaction produced considerable detachment by cohesive failure of the membrane. In this study, we examined the hypothesis that reducing the SA-biotin bond affinity could reduce cohesive failure without reducing overall cell detachment. Two mutants of SA, W120F and W120A in which the tryptophan residue at position 120 of the SA molecule was substituted by phenylalanine and alanine, respectively, were characterized and tested in cell adhesion experiments. The binding affinity (K(A)) of SA to adsorbed biotin-labeled bovine serum albumin (b-BSA) ranged from 5.2+/-0.1 x 10(10)M(-1) for wild-type to 3.3+/-0.2 x 10(9)M(-1) for W120F and 4.1+/-1.0 x 10(6)M(-1) for W120A. One hour after cell attachment, the critical shear stress was 26.8+/-2.9 dyn/cm(2) for WT, 26.6+/-3.0 dyn/cm(2) for W120F, and 15.4+/-3.0 dyn/cm(2) for W120A. The focal contact areas of adherent cells were greater for the WT and W120F than the lower affinity mutant, W120A. When shear flow was applied to detach adherent cells, adhesive failure (ligand bond breakage) was favored over cohesive failure (membrane rupture), as the SA binding affinity decreased. Thus, cell adhesion augmented by SA-biotin linkages is dependent on the affinity constants of the SA-biotin bonds, but the reduction in cohesive failure was offset by a reduced strength of adhesion.

Duke Scholars

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

February 2003

Volume

24

Issue

4

Start / End Page

559 / 570

Related Subject Headings

  • Streptavidin
  • Protein Binding
  • Mathematics
  • Ligands
  • Integrins
  • Endothelium, Vascular
  • Cell Membrane
  • Cell Adhesion
  • Cattle
  • Biotin
 

Citation

APA
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ICMJE
MLA
NLM
Chan, B. P., Chilkoti, A., Reichert, W. M., & Truskey, G. A. (2003). Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells. Biomaterials, 24(4), 559–570. https://doi.org/10.1016/s0142-9612(02)00367-8
Chan, Bernard P., Ashutosh Chilkoti, William M. Reichert, and George A. Truskey. “Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells.Biomaterials 24, no. 4 (February 2003): 559–70. https://doi.org/10.1016/s0142-9612(02)00367-8.
Chan BP, Chilkoti A, Reichert WM, Truskey GA. Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells. Biomaterials. 2003 Feb;24(4):559–70.
Chan, Bernard P., et al. “Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells.Biomaterials, vol. 24, no. 4, Feb. 2003, pp. 559–70. Epmc, doi:10.1016/s0142-9612(02)00367-8.
Chan BP, Chilkoti A, Reichert WM, Truskey GA. Effect of streptavidin affinity mutants on the integrin-independent adhesion of biotinylated endothelial cells. Biomaterials. 2003 Feb;24(4):559–570.
Journal cover image

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

February 2003

Volume

24

Issue

4

Start / End Page

559 / 570

Related Subject Headings

  • Streptavidin
  • Protein Binding
  • Mathematics
  • Ligands
  • Integrins
  • Endothelium, Vascular
  • Cell Membrane
  • Cell Adhesion
  • Cattle
  • Biotin