Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 May 7;12:RP88991. doi: 10.7554/eLife.88991

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2023, Duewell, Wilson et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

PMC Copyright notice

Figure 2. — (A) Cartoon schematic showing conjugation of pY peptides (+/- Alexa488 label) using thiol-reactive maleimide lipids (MCC-PE). (B) Representative image showing the single molecule localization of Alexa488-pY. Particle detection (purple circles) was used to quantify the number of pY peptides per µm². (C) Relationship between the total pY solution concentration (x-axis) used for covalent conjugation, the bulk membrane intensity of covalently attached Alexa488-pY (left y-axis), and the final surface density of pY peptides per µm² (right y-axis). (D) Relationship between the total pY solution conjugation concentration and bulk membrane intensity of measured in the presence of 50 nM nSH2-Cy3. (E–G) Membrane localization dynamics of Dy647-PI3Kβ measured on supported lipid bilayers (SLBs) containing a range of pY surface densities (250–15,000 pY/µm², based on Figure 1C). (E) Bulk membrane localization of 10 nM Dy647-PI3Kβ as a function of pY density. (F) Single molecule dwell time distributions measured in the presence of 5 pM Dy647-PI3Kβ. Data plotted as log₁₀(1–CDF) (cumulative distribution frequency). (G) Step size distributions showing Dy647-PI3Kβ single molecule displacements from >500 particles (>10,000 steps) per pY surface density. (H–J) Membrane localization dynamics of Dy647-PI3Kβ nSH2* (R358A) and cSH2* (R649A) mutants measured on SLBs containing ~15,000 pY/µm² (10 µM conjugation concentration). (H) Bulk membrane absorption kinetics of 10 nM Dy647-PI3Kβ (WT, nSH2*, and cSH2*). (I) Single molecule dwell time distributions measured in the presence of 5 pM Dy647-PI3Kβ (WT, nSH2*, and cSH2*). Data plotted as log₁₀(1–CDF) (cumulative distribution frequency). (J) Step size distributions showing single molecule displacements of >500 particles (>10,000 steps) in the presence of 5 pM Dy647-PI3Kβ (WT, nSH2*, and cSH2*). Membrane composition: 96% DOPC, 2% PI(4,5)P₂, 2% MCC-PE.

Figure 2—source data 1. Related to .Figure 2C.

elife-88991-fig2-data1.xlsx^{(9.2KB, xlsx)}

Figure 2—source data 2. Related to .Figure 2D.

elife-88991-fig2-data2.xlsx^{(9.3KB, xlsx)}

Figure 2—source data 3. Related to .Figure 2E.

elife-88991-fig2-data3.xlsx^{(16.7KB, xlsx)}

Figure 2—source data 4. Related to .Figure 2F.

elife-88991-fig2-data4.xlsx^{(54KB, xlsx)}

Figure 2—source data 5. Related to .Figure 2G.

elife-88991-fig2-data5.xlsx^{(1,014.5KB, xlsx)}

Figure 2—source data 6. Related to .Figure 2H.

elife-88991-fig2-data6.xlsx^{(14.3KB, xlsx)}

Figure 2—source data 7. Related to .Figure 2I.

elife-88991-fig2-data7.xlsx^{(46KB, xlsx)}

Figure 2—source data 8. Related to .Figure 2J.

elife-88991-fig2-data8.xlsx^{(550.5KB, xlsx)}