Direct Observation of Dynamic Protein Interactions Involving Human Microtubules Using Solid-state NMR Spectroscopy

Overview

Journal Nat Commun

Specialty Biology

Date 2020 Jan 4

PMID 31896752

Citations 8

Authors

Yanzhang Luo

ShengQi Xiang

Peter Jan Hooikaas

Laura van Bezouwen

A S Jijumon

Carsten Janke

Friedrich Forster

Anna Akhmanova

Marc Baldus

Affiliations

Soon will be listed here.

Abstract

Microtubules are important components of the eukaryotic cytoskeleton. Their structural organization is regulated by nucleotide binding and many microtubule-associated proteins (MAPs). While cryo-EM and X-ray crystallography have provided detailed views of interactions between MAPs with the microtubule lattice, little is known about how MAPs and their intrinsically disordered regions interact with the dynamic microtubule surface. NMR carries the potential to directly probe such interactions but so far has been precluded by the low tubulin yield. We present a protocol to produce [C, N]-labeled, functional microtubules (MTs) from human cells for solid-state NMR studies. This approach allowed us to demonstrate that MAPs can differently modulate the fast time-scale dynamics of C-terminal tubulin tails, suggesting distinct interaction modes. Our results pave the way for in-depth NMR studies of protein dynamics involved in MT assembly and their interactions with other cellular components.

Citing Articles

Elucidating the Subcellular Localization of GLRaV-3 Proteins Encoded by the Unique Gene Block in Suggests Implications on Plant Host Suppression.

Lameront P, Shabanian M, Currie L, Fust C, Li C, Clews A Biomolecules. 2024; 14(8).

PMID: 39199365 PMC: 11352578. DOI: 10.3390/biom14080977.

A structural and dynamic visualization of the interaction between MAP7 and microtubules.

Adler A, Bangera M, Beugelink J, Bahri S, Ingen H, Moores C Nat Commun. 2024; 15(1):1948.

PMID: 38431715 PMC: 10908866. DOI: 10.1038/s41467-024-46260-5.

High Expression of Microtubule-associated Protein TBCB Predicts Adverse Outcome and Immunosuppression in Acute Myeloid Leukemia.

Wang B, Wang W, Li Q, Guo T, Yang S, Shi J J Cancer. 2023; 14(10):1707-1724.

PMID: 37476188 PMC: 10355208. DOI: 10.7150/jca.84215.

H-detected characterization of carbon-carbon networks in highly flexible protonated biomolecules using MAS NMR.

Bahri S, Safeer A, Adler A, Smedes H, Ingen H, Baldus M J Biomol NMR. 2023; 77(3):111-119.

PMID: 37289305 PMC: 10307723. DOI: 10.1007/s10858-023-00415-6.

Resonance assignments of the microtubule-binding domain of the microtubule-associated protein 7 (MAP7).

Adler A, Kjaer L, Beugelink J, Baldus M, Ingen H Biomol NMR Assign. 2023; 17(1):83-88.

PMID: 37099260 PMC: 10232616. DOI: 10.1007/s12104-023-10124-8.

References

Tan R, Lam A, Tan T, Han J, Nowakowski D, Vershinin M . Microtubules gate tau condensation to spatially regulate microtubule functions. Nat Cell Biol. 2019; 21(9):1078-1085. PMC: 6748660. DOI: 10.1038/s41556-019-0375-5. View

Fung B, Khitrin A, Ermolaev K . An improved broadband decoupling sequence for liquid crystals and solids. J Magn Reson. 2000; 142(1):97-101. DOI: 10.1006/jmre.1999.1896. View

Chaudhary A, Lu H, Krementsova E, Bookwalter C, Trybus K, Hendricks A . MAP7 regulates organelle transport by recruiting kinesin-1 to microtubules. J Biol Chem. 2019; 294(26):10160-10171. PMC: 6664170. DOI: 10.1074/jbc.RA119.008052. View

Lakomek N, Yavuz H, Jahn R, Perez-Lara A . Structural dynamics and transient lipid binding of synaptobrevin-2 tune SNARE assembly and membrane fusion. Proc Natl Acad Sci U S A. 2019; 116(18):8699-8708. PMC: 6500178. DOI: 10.1073/pnas.1813194116. View

Nogales E, Zhang R . Visualizing microtubule structural transitions and interactions with associated proteins. Curr Opin Struct Biol. 2016; 37:90-6. PMC: 4834239. DOI: 10.1016/j.sbi.2015.12.009. View

Hernandez-Vega A, Braun M, Scharrel L, Jahnel M, Wegmann S, Hyman B . Local Nucleation of Microtubule Bundles through Tubulin Concentration into a Condensed Tau Phase. Cell Rep. 2017; 20(10):2304-2312. PMC: 5828996. DOI: 10.1016/j.celrep.2017.08.042. View

Lefevre J, Chernov K, Joshi V, Delga S, Toma F, Pastre D . The C terminus of tubulin, a versatile partner for cationic molecules: binding of Tau, polyamines, and calcium. J Biol Chem. 2010; 286(4):3065-78. PMC: 3024800. DOI: 10.1074/jbc.M110.144089. View

Pan X, Cao Y, Stucchi R, Hooikaas P, Portegies S, Will L . MAP7D2 Localizes to the Proximal Axon and Locally Promotes Kinesin-1-Mediated Cargo Transport into the Axon. Cell Rep. 2019; 26(8):1988-1999.e6. PMC: 6381606. DOI: 10.1016/j.celrep.2019.01.084. View

Hooikaas P, Martin M, Muhlethaler T, Kuijntjes G, Peeters C, Katrukha E . MAP7 family proteins regulate kinesin-1 recruitment and activation. J Cell Biol. 2019; 218(4):1298-1318. PMC: 6446838. DOI: 10.1083/jcb.201808065. View

10.

Guedes-Dias P, Nirschl J, Abreu N, Tokito M, Janke C, Magiera M . Kinesin-3 Responds to Local Microtubule Dynamics to Target Synaptic Cargo Delivery to the Presynapse. Curr Biol. 2019; 29(2):268-282.e8. PMC: 6342647. DOI: 10.1016/j.cub.2018.11.065. View

11.

Spoerner M, Karl M, Lopes P, Hoering M, Loeffel K, Nuehs A . High pressure P NMR spectroscopy on guanine nucleotides. J Biomol NMR. 2016; 67(1):1-13. DOI: 10.1007/s10858-016-0079-0. View

12.

Etzkorn M, Bockmann A, Lange A, Baldus M . Probing molecular interfaces using 2D magic-angle-spinning NMR on protein mixtures with different uniform labeling. J Am Chem Soc. 2004; 126(45):14746-51. DOI: 10.1021/ja0479181. View

13.

Kubo S, Nishida N, Udagawa Y, Takarada O, Ogino S, Shimada I . A gel-encapsulated bioreactor system for NMR studies of protein-protein interactions in living mammalian cells. Angew Chem Int Ed Engl. 2012; 52(4):1208-11. DOI: 10.1002/anie.201207243. View

14.

Narasimhan S, Mance D, Pinto C, Weingarth M, Bonvin A, Baldus M . Rapid Prediction of Multi-dimensional NMR Data Sets Using FANDAS. Methods Mol Biol. 2017; 1688:111-132. DOI: 10.1007/978-1-4939-7386-6_6. View

15.

Atherton J, Luo Y, Xiang S, Yang C, Rai A, Jiang K . Structural determinants of microtubule minus end preference in CAMSAP CKK domains. Nat Commun. 2019; 10(1):5236. PMC: 6868217. DOI: 10.1038/s41467-019-13247-6. View

16.

Peng J . Exposing the Moving Parts of Proteins with NMR Spectroscopy. J Phys Chem Lett. 2012; 3(8):1039-1051. PMC: 3336369. DOI: 10.1021/jz3002103. View

17.

Yan S, Guo C, Hou G, Zhang H, Lu X, Williams J . Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy. Proc Natl Acad Sci U S A. 2015; 112(47):14611-6. PMC: 4664305. DOI: 10.1073/pnas.1509852112. View

18.

Xiang S, le Paige U, Horn V, Houben K, Baldus M, Ingen H . Site-Specific Studies of Nucleosome Interactions by Solid-State NMR Spectroscopy. Angew Chem Int Ed Engl. 2018; 57(17):4571-4575. PMC: 5947581. DOI: 10.1002/anie.201713158. View

19.

Alam T, Holland G . (1)H-(13)C INEPT MAS NMR correlation experiments with (1)H-(1)H mediated magnetization exchange to probe organization in lipid biomembranes. J Magn Reson. 2006; 180(2):210-21. DOI: 10.1016/j.jmr.2006.02.013. View

20.

Weaver B . How Taxol/paclitaxel kills cancer cells. Mol Biol Cell. 2014; 25(18):2677-81. PMC: 4161504. DOI: 10.1091/mbc.E14-04-0916. View