Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers

Chia Fen Hsieh, Bo Jui Chang, Chyi-Huey Bai, Hsuan Yi Chen, Sien Chi, Long Hsu, Jin Wu Tsai, Chi Hung Lin

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Integrin receptors serve as both mechanical links and signal transduction mediators between the cell and its environment. Experimental evidence demonstrates that conformational changes and lateral clustering of the integrin proteins may affect their binding to ligands and regulate downstream cellular responses; however, experimental links between the structural and functional correlations of the ligand-receptor interactions are not yet elucidated. In the present report, we utilized optical tweezers to measure the dynamic binding between the snake venom rhodostomin, coated on a microparticle and functioned as a ligand, and the membrane receptor integrin αIIbβ 3 expressed on a Chinese Hamster Ovary (CHO) cell. A progressive increase of total binding affinity was found between the bead and CHO cell in the first 300 sec following optical tweezers-guided contact. Further analysis of the cumulative data revealed the presence of "unit binding force" presumably exerted by a single rhodostomin-integrin pair. Interestingly, two such units were found. Among the measurements of less total binding forces, presumably taken at the early stage of ligand-receptor interactions, a unit of 4.15 pN per molecule pair was derived. This unit force dropped to 2.54 pN per molecule pair toward the later stage of interactions when the total binding forces were relatively large. This stepped change of single molecule pair binding affinity was not found when mutant rhodostomin proteins were used as ligands (a single unit of 1.81 pN per pair was found). These results were interpreted along with the current knowledge about the conformational changes of integrins during the "molecule activation" process.

Original languageEnglish
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsK. Dholakia, G.C. Spalding
Pages215-224
Number of pages10
Volume5514
DOIs
Publication statusPublished - 2004
Externally publishedYes
EventOptical Trapping and Optical Micromanipulation - Denver, CO, United States
Duration: Aug 2 2004Aug 6 2004

Other

OtherOptical Trapping and Optical Micromanipulation
CountryUnited States
CityDenver, CO
Period8/2/048/6/04

Fingerprint

Optical tweezers
affinity
Ligands
Cells
Molecules
ligands
interactions
ovaries
hamsters
Proteins
Signal transduction
molecules
proteins
snakes
microparticles
Chemical activation
cells
beads
Membranes
activation

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Hsieh, C. F., Chang, B. J., Bai, C-H., Chen, H. Y., Chi, S., Hsu, L., ... Lin, C. H. (2004). Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers. In K. Dholakia, & G. C. Spalding (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 5514, pp. 215-224). [27] https://doi.org/10.1117/12.558653

Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers. / Hsieh, Chia Fen; Chang, Bo Jui; Bai, Chyi-Huey; Chen, Hsuan Yi; Chi, Sien; Hsu, Long; Tsai, Jin Wu; Lin, Chi Hung.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / K. Dholakia; G.C. Spalding. Vol. 5514 2004. p. 215-224 27.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Hsieh, CF, Chang, BJ, Bai, C-H, Chen, HY, Chi, S, Hsu, L, Tsai, JW & Lin, CH 2004, Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers. in K Dholakia & GC Spalding (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 5514, 27, pp. 215-224, Optical Trapping and Optical Micromanipulation, Denver, CO, United States, 8/2/04. https://doi.org/10.1117/12.558653
Hsieh CF, Chang BJ, Bai C-H, Chen HY, Chi S, Hsu L et al. Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers. In Dholakia K, Spalding GC, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 5514. 2004. p. 215-224. 27 https://doi.org/10.1117/12.558653
Hsieh, Chia Fen ; Chang, Bo Jui ; Bai, Chyi-Huey ; Chen, Hsuan Yi ; Chi, Sien ; Hsu, Long ; Tsai, Jin Wu ; Lin, Chi Hung. / Identification of stepped changes of binding affinity during interactions between the disintegrin rhodostomin and integrin α IIbβ3 in living cells using optical tweezers. Proceedings of SPIE - The International Society for Optical Engineering. editor / K. Dholakia ; G.C. Spalding. Vol. 5514 2004. pp. 215-224
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