A correlated force-optical study on the self-assembly behavior of annexin V on model membranes: effect of dye conjugation.
We have examined the self-assembled membrane-bound aggregates of two annexin V (A5) dye conjugates and compared them to those from native A5. Native A5 and FITC-labeled A5 (A5-FITC) both formed discrete well-defined crystalline monolayer domains of p6 symmetry. However, A5-FITC also showed additional domains with a corrugated appearance not observed in native A5. In contrast, Cy3-labeled A5 (A5-Cy3) showed a mixture of crystalline monolayer and irregular multilayered domains, with the ratio of the two types varying significantly from sample to sample, and also required a much longer incubation time than native A5 and A5-FITC. When A5-FITC and A5-Cy3 were co-incubated on the same bilayer, well-defined crystalline monolayer domains containing both A5-FITC and A5-Cy3 were consistently observed at a much shorter incubation time than that of pure A5-Cy3 alone, indicating that A5-FITC facilitates the inclusion of A5-Cy3. These results suggest that dye labels can affect A5 2D self-assembly and crystal formation on membrane surfaces.
Han
Jason J
JJ
Department of Chemistry, and Department of Physics, Yonsei University, Seoul, Korea.
Park
Seung-Han
SH
Boo
Doo Wan
DW
D017304:Annexin A5 / Q000737:chemistry / Q000378:metabolism*; D002232:Carbocyanines / Q000378:metabolism / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism / Q000494:pharmacology; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism* / Q000494:pharmacology*; D007185:Incubators; D008051:Lipid Bilayers / Q000378:metabolism*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D055095:Optics and Photonics; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D013696:Temperature
D017304:Annexin A5; D002232:Carbocyanines; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008567:Membranes, Artificial; C089187:cyanine dye 3; D016650:Fluorescein-5-isothiocyanate