The Fluid Membrane determines Mechanics of Red Blood Cell Extracellular Vesicles and is Softened in Hereditary Spherocytosis

Vorselen, Daan; Dommelen, Susan M. van; Sorkin, Raya; Schiller, Jürgen; Wijk, Richard van; Schiffelers, Raymond M.; Wuite, Gijs J.L.; Roos, Wouter H.

doi:10.1101/212456

Cited by 5 publications

(10 citation statements)

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“…One striking observation in our study is the similarity in the mechanics of vesicles from different cells and origins, as well as isolation methods, blood donors, and experimentalists performing the vesicle isolation. We find that bending modulus values obtained here for RT incubated RBC are very comparable to those we previously measured in a different set of experiments with vesicles produced at similar conditions of RT 20 h incubation, [16] demonstrating again the robustness of our method.…”

Section: Discussionsupporting

confidence: 87%

“…Very few previous pioneering studies addressed the importance of mechanical properties of extracellular vesicles and so far the pressurization of vesicles due to surface interactions has not been taken into account. Our recently developed approach does take pressurization into account. It allows reliable comparison of different vesicle populations, as we are able to estimate the bending modulus of the vesicles, an intrinsic material property that is independent of the measuring conditions.…”

Section: Discussionmentioning

confidence: 99%

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Nanomechanics of Extracellular Vesicles Reveals Vesiculation Pathways

Sorkin

Huisjes

Bošković

et al. 2018

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Extracellular vesicles (EVs) are emerging as important mediators of cell-cell communication as well as potential disease biomarkers and drug delivery vehicles. However, the mechanical properties of these vesicles are largely unknown, and processes leading to microvesicle-shedding from the plasma membrane are not well understood. Here an in depth atomic force microscopy force spectroscopy study of the mechanical properties of natural EVs is presented. It is found that several natural vesicles of different origin have a different composition of lipids and proteins, but similar mechanical properties. However, vesicles generated by red blood cells (RBC) at different temperatures/incubation times are different mechanically. Quantifying the lipid content of EVs reveals that their stiffness decreases with the increase in their protein/lipid ratio. Further, by maintaining RBC at "extreme" nonphysiological conditions, the cells are pushed to utilize different vesicle generation pathways. It is found that RBCs can generate protein-rich soft vesicles, possibly driven by protein aggregation, and low membrane-protein content stiff vesicles, likely driven by cytoskeleton-induced buckling. Since similar cortical cytoskeleton to that of the RBC exists on the membranes of most mammalian cells, our findings help advancing the understanding of the fundamental process of vesicle generation.

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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Nanomechanics of Extracellular Vesicles Reveals Vesiculation Pathways

Sorkin

Huisjes

Bošković

et al. 2018

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“…The effect of proteins on the membrane mechanics has been investigated in various studies, some of which reported a significant increase in bending modulus of vesicles 50,51 , while others have shown either a neutral effect 52 or a decrease in the membrane stiffness. [53][54][55][56] Generally speaking, short peptides usually decrease the rigidity (stiffness) of the membrane 54 , while the effect of larger membrane proteins could be either neutral 52 or decreasing. 53,55 The reason for this is the hydrophobic mismatch due to the differences in the thickness of hydrophobic chain of the lipid molecule and the length of the hydrophobic surface of the protein 57 that causes a local bilayer thinning in the case of a short peptide.…”

Section: Effect Of Other Moleculesmentioning

confidence: 99%

“…[53][54][55][56] Generally speaking, short peptides usually decrease the rigidity (stiffness) of the membrane 54 , while the effect of larger membrane proteins could be either neutral 52 or decreasing. 53,55 The reason for this is the hydrophobic mismatch due to the differences in the thickness of hydrophobic chain of the lipid molecule and the length of the hydrophobic surface of the protein 57 that causes a local bilayer thinning in the case of a short peptide. The effect of proteins is discussed in more details in Chapter 4. mineral processing and biological analysis.…”

Section: Effect Of Other Moleculesmentioning

confidence: 99%

“…186 The differentiation, however, has been mainly dictated by the membrane lipidome, while for medically relevant vesicles proteins embedded in the membrane play a major role. 55 In fact, membrane proteins have been recently suggested to be the main contributor to the stiffness of natural EVs. 56 The over and underexpression of certain proteins in extracellular vesicles has been observed in many physiological and pathological conditions; however, a simple method to sort vesicles based on contrast in protein content is yet to be developed.…”

Section: Introductionmentioning

confidence: 99%

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The Effect of Proteome and Lipidome on the Behavior of Membrane Bound Systems in Thermally-Assisted Acoustophoresis

Mirtaheri¹

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I would like to express my gratitude to all the people who have helped me and given me support throughout the completion of my PhD studies. First, to my parents, Mehran and Mohammad, who have always loved and supported me. Of course, I could not have done this without the support and encouragement of other family members including my brother Ali. I would like to thank my advisor, Dr. Bilal El-Zahab, for guiding me and helping me reach my potential. His technical guidance, support, and encouragement were very important factors in my completing this effort. This dissertation would not have been possible without the support of my committee members Dr. Shekhar Bhansali, Dr. Cesar Levy, Dr. Ibrahim Tansel and Dr. Yiding Cao all of whom deserve special thanks for providing support, professionally and personally, along the way. Shervin, Maedeh, Azadeh, Amirali and Arezou I considered whom my best friends at FIU, deserve a special mention. I am indebted to you both for your support and friendship over these years. A very special acknowledgement goes to Ata, who encouraged me and given me unwavering support from the very first days of my Ph.D. studies and even before, and constantly made me feel like I am not alone. Finally, I would like to acknowledge the teachers, professors, and other professionals who either directly trained me, or inspired me in some way. vi

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Thermally Assisted Acoustofluidic Separation Based on Membrane Protein Content

2019

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The over- and under-expression of certain proteins in extracellular vesicles has been observed in many physiological and pathological conditions; however, a simple method to sort vesicles based on contrast in protein content is yet to be developed. We herein present a nonaffinity-based method for rapid and inexpensive isolation of lipid vesicles based on their membrane protein content. Based on a composition-specific thermophysical property change of vesicles at different protein contents, an acoustic property change that enabled an acoustophoretic separation was observed. This change was demonstrated in a thermally modulated acoustofluidic device in the form of a shift in vesicle migration from the nodal plane to antinodal plane at a specific temperature known as the acoustic contrast temperature (T Φ). Using phosphatidylcholine vesicles containing the membrane proteins gramicidin D, alamethicin, and melittin at molar contents ranging from 0.001% to 10%, we observed that increasing the membrane protein content brought about conformational changes in the membrane which afforded the vesicles distinctive acoustic properties. Then, by establishing an acoustic contrast temperature window, vesicles with the same protein but different molar content were successfully separated. The efficiency of the separation was studied for various vesicle mixtures and a separation efficiency as high as 97% was accomplished. In order to confirm the technique’s applicability for biological samples, sheep red blood cells with various melittin peptide contents similarly demonstrated the depressing effects of melittin on membrane bending modulus and depressed the T Φ of the cells. This method holds promise for a myriad of applications in the biomedical field, especially in bioanalytical research.

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The Fluid Membrane determines Mechanics of Red Blood Cell Extracellular Vesicles and is Softened in Hereditary Spherocytosis

Abstract: was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

Cited by 5 publications

References 71 publications

Nanomechanics of Extracellular Vesicles Reveals Vesiculation Pathways

Nanomechanics of Extracellular Vesicles Reveals Vesiculation Pathways

The Effect of Proteome and Lipidome on the Behavior of Membrane Bound Systems in Thermally-Assisted Acoustophoresis

Thermally Assisted Acoustofluidic Separation Based on Membrane Protein Content

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