Supramolecular Assembly in Live Cells Mapped by Real-Time Phasor-Fluorescence Lifetime Imaging

Overview

Journal J Am Chem Soc

Publisher American Chemical Society

Specialty Chemistry

Date 2024 Apr 19

PMID 38639465

Authors

Yong Ren

Zhixuan Zhou

Konrad Maxeiner

Anke Kaltbeitzel

Iain Harley

Jiaqi Xing

Yingke Wu

Manfred Wagner

Katharina Landfester

Ingo Lieberwirth

Tanja Weil

David Y W Ng

Affiliations

Soon will be listed here.

Abstract

The complex dynamics and transience of assembly pathways in living systems complicate the understanding of these molecular to nanoscale processes. Current technologies are unable to track the molecular events leading to the onset of assembly, where real-time information is imperative to correlate their rich biology. Using a chemically designed pro-assembling molecule, we map its transformation into nanofibers and their fusion with endosomes to form hollow fiber clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) in epithelial cells (L929, A549, MDA-MB 231) and correlative light-electron microscopy and tomography (CLEM), spatiotemporal splicing of the assembly events shows time-correlated metabolic dysfunction. The biological impact begins with assembly-induced endosomal disruption that reduces glucose transport into the cells, which, in turn, stymies mitochondrial respiration.

Citing Articles

Precise Regulations at the Subcellular Level through Intracellular Polymerization, Assembly, and Transformation.

He L, Meng F, Chen R, Qin J, Sun M, Fan Z JACS Au. 2024; 4(11):4162-4186.

PMID: 39610726 PMC: 11600172. DOI: 10.1021/jacsau.4c00849.

References

Qualls-Histed S, Nielsen C, MacGurn J . Lysosomal trafficking of the glucose transporter GLUT1 requires sequential regulation by TXNIP and ubiquitin. iScience. 2023; 26(3):106150. PMC: 9986520. DOI: 10.1016/j.isci.2023.106150. View

Sarkar A, Kolsch J, Berac C, Venugopal A, Sasmal R, Otter R . Impact of NDI-Core Substitution on the pH-Responsive Nature of Peptide-Tethered Luminescent Supramolecular Polymers. ChemistryOpen. 2020; 9(3):346-350. PMC: 7080532. DOI: 10.1002/open.202000017. View

Fleming S, Ulijn R . Design of nanostructures based on aromatic peptide amphiphiles. Chem Soc Rev. 2014; 43(23):8150-77. DOI: 10.1039/c4cs00247d. View

Bowerman C, Nilsson B . Self-assembly of amphipathic β-sheet peptides: insights and applications. Biopolymers. 2012; 98(3):169-84. DOI: 10.1002/bip.22058. View

Stolowitz M, Ahlem C, Hughes K, KAISER R, Kesicki E, Li G . Phenylboronic acid-salicylhydroxamic acid bioconjugates. 1. A novel boronic acid complex for protein immobilization. Bioconjug Chem. 2001; 12(2):229-39. DOI: 10.1021/bc0000942. View

Wang N, Zhang S, Yuan Y, Xu H, Defossa E, Matter H . Molecular basis for inhibiting human glucose transporters by exofacial inhibitors. Nat Commun. 2022; 13(1):2632. PMC: 9098912. DOI: 10.1038/s41467-022-30326-3. View

Chagri S, Ng D, Weil T . Designing bioresponsive nanomaterials for intracellular self-assembly. Nat Rev Chem. 2023; 6(5):320-338. PMC: 8972907. DOI: 10.1038/s41570-022-00373-x. View

Wang T, Menard-Moyon C, Bianco A . Self-assembly of amphiphilic amino acid derivatives for biomedical applications. Chem Soc Rev. 2022; 51(9):3535-3560. DOI: 10.1039/d1cs01064f. View

Shinde S, Maddika S . PTEN Regulates Glucose Transporter Recycling by Impairing SNX27 Retromer Assembly. Cell Rep. 2017; 21(6):1655-1666. PMC: 5695913. DOI: 10.1016/j.celrep.2017.10.053. View

10.

Ulery P, Beers J, Mikhailenko I, Tanzi R, Rebeck G, Hyman B . Modulation of beta-amyloid precursor protein processing by the low density lipoprotein receptor-related protein (LRP). Evidence that LRP contributes to the pathogenesis of Alzheimer's disease. J Biol Chem. 2000; 275(10):7410-5. DOI: 10.1074/jbc.275.10.7410. View

11.

Ling D, Song H, Garza D, Neufeld T, Salvaterra P . Abeta42-induced neurodegeneration via an age-dependent autophagic-lysosomal injury in Drosophila. PLoS One. 2009; 4(1):e4201. PMC: 2626277. DOI: 10.1371/journal.pone.0004201. View

12.

Marchetti P, Fovez Q, Germain N, Khamari R, Kluza J . Mitochondrial spare respiratory capacity: Mechanisms, regulation, and significance in non-transformed and cancer cells. FASEB J. 2020; 34(10):13106-13124. DOI: 10.1096/fj.202000767R. View

13.

Mishra A, Lai G, Schmidt N, Sun V, Rodriguez A, Tong R . Translocation of HIV TAT peptide and analogues induced by multiplexed membrane and cytoskeletal interactions. Proc Natl Acad Sci U S A. 2011; 108(41):16883-8. PMC: 3193208. DOI: 10.1073/pnas.1108795108. View

14.

de Boer P, Hoogenboom J, Giepmans B . Correlated light and electron microscopy: ultrastructure lights up!. Nat Methods. 2015; 12(6):503-13. DOI: 10.1038/nmeth.3400. View

15.

Di Lisa F, Menabo R, Canton M, Barile M, Bernardi P . Opening of the mitochondrial permeability transition pore causes depletion of mitochondrial and cytosolic NAD+ and is a causative event in the death of myocytes in postischemic reperfusion of the heart. J Biol Chem. 2000; 276(4):2571-5. DOI: 10.1074/jbc.M006825200. View

16.

Kang D, Pietrzik C, Baum L, Chevallier N, Merriam D, Kounnas M . Modulation of amyloid beta-protein clearance and Alzheimer's disease susceptibility by the LDL receptor-related protein pathway. J Clin Invest. 2000; 106(9):1159-66. PMC: 301422. DOI: 10.1172/JCI11013. View

17.

Wang L, Tran M, DEste E, Roberti J, Koch B, Xue L . A general strategy to develop cell permeable and fluorogenic probes for multicolour nanoscopy. Nat Chem. 2019; 12(2):165-172. DOI: 10.1038/s41557-019-0371-1. View

18.

Frederix P, Ulijn R, Hunt N, Tuttle T . Virtual Screening for Dipeptide Aggregation: Toward Predictive Tools for Peptide Self-Assembly. J Phys Chem Lett. 2013; 2(19):2380-2384. PMC: 3688361. DOI: 10.1021/jz2010573. View

19.

Micsonai A, Wien F, Kernya L, Lee Y, Goto Y, Refregiers M . Accurate secondary structure prediction and fold recognition for circular dichroism spectroscopy. Proc Natl Acad Sci U S A. 2015; 112(24):E3095-103. PMC: 4475991. DOI: 10.1073/pnas.1500851112. View

20.

Schmid S, Abbel R, Schenning A, Meijer E, Herz L . Energy transfer processes along a supramolecular chain of π-conjugated molecules. Philos Trans A Math Phys Eng Sci. 2012; 370(1972):3787-801. DOI: 10.1098/rsta.2011.0205. View