Elucidation of transient protein-protein interactions within carrier protein-dependent biosynthesis.

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / metabolism Acetyltransferases / metabolism Acyl Carrier Protein / metabolism Alcohol Oxidoreductases / metabolism Binding Sites Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) / metabolism Escherichia coli / enzymology Escherichia coli Proteins / metabolism Fatty Acid Synthase, Type II / metabolism Fatty Acids / biosynthesis Lysophospholipase / metabolism Molecular Docking Simulation Periplasmic Proteins / metabolism Protein Binding Protein Interaction Domains and Motifs Proton Magnetic Resonance Spectroscopy

Journal

Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179

Informations de publication

Date de publication:
16 03 2021
Historique:
received: 15 09 2020
accepted: 11 02 2021
entrez: 17 3 2021
pubmed: 18 3 2021
medline: 11 8 2021
Statut: epublish

Résumé

Fatty acid biosynthesis (FAB) is an essential and highly conserved metabolic pathway. In bacteria, this process is mediated by an elaborate network of protein•protein interactions (PPIs) involving a small, dynamic acyl carrier protein that interacts with dozens of other partner proteins (PPs). These PPIs have remained poorly characterized due to their dynamic and transient nature. Using a combination of solution-phase NMR spectroscopy and protein-protein docking simulations, we report a comprehensive residue-by-residue comparison of the PPIs formed during FAB in Escherichia coli. This technique describes and compares the molecular basis of six discrete binding events responsible for E. coli FAB and offers insights into a method to characterize these events and those in related carrier protein-dependent pathways.

Identifiants

DOI: 10.1038/s42003-021-01838-3 PMID: 33727677 PMC: PMC7966745
pubmed: 33727677
doi: 10.1038/s42003-021-01838-3
pii: 10.1038/s42003-021-01838-3
pmc: PMC7966745
doi:

Substances chimiques

Acyl Carrier Protein 0
Escherichia coli Proteins 0
Fatty Acids 0
Periplasmic Proteins 0
acpP protein, E coli 0
Alcohol Oxidoreductases EC 1.1.-
acetoacetyl-CoA reductase EC 1.1.1.36
Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) EC 1.3.1.9
fabI protein, E coli EC 1.3.1.9
Acetyltransferases EC 2.3.1.-
fabF protein, E coli EC 2.3.1.179
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase EC 2.3.1.41
FabB protein, E coli EC 2.3.1.41
Lysophospholipase EC 3.1.1.5
tesA protein, E coli EC 3.1.2.-
Fatty Acid Synthase, Type II EC 6.-

Types de publication

Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

340

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM095970
Pays : United States
Organisme : NIAID NIH HHS
ID : R21 AI156484
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM131881
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM112584
Pays : United States

Références

Crosby, J. & Crump, M. P. The structural role of the carrier protein—active controller or passive carrier. Nat. Prod. Rep. 29, 1111–1137 (2012).
pubmed: 22930263 doi: 10.1039/c2np20062g
Lai, J. R., Koglin, A. & Walsh, C. T. Carrier protein structure and recognition in polyketide and nonribosomal peptide biosynthesis. Biochemistry 45, 14869–14879 (2006).
pubmed: 17154525 doi: 10.1021/bi061979p
Schweizer, E. & Hofmann, J. Microbial type I fatty acid synthases (FAS): major players in a network of cellular FAS systems. Microbiol. Mol. Biol. Rev. 68, 501–517 (2004).
pubmed: 15353567 pmcid: 515254 doi: 10.1128/MMBR.68.3.501-517.2004
Gully, D. & Bouveret, E. A protein network for phospholipid synthesis uncovered by a variant of the tandem affinity purification method in Escherichia coli. Proteomics 6, 282–293 (2006).
pubmed: 16294310 doi: 10.1002/pmic.200500115
De Lay, N. R. & Cronan, J. E. In vivo functional analyses of the type II acyl carrier proteins of fatty acid biosynthesis. J. Biol. Chem. 282, 20319–20328 (2007).
pubmed: 17522044 doi: 10.1074/jbc.M703789200
Gully, D., Moinier, D., Loiseau, L. & Bouveret, E. New partners of acyl carrier protein detected in Escherichia coli by tandem affinity purification. FEBS Lett. 548, 90–96 (2003).
pubmed: 12885413 doi: 10.1016/S0014-5793(03)00746-4
Roujeinikova, A. et al. Structural studies of fatty acyl-(acyl carrier protein) thioesters reveal a hydrophobic binding cavity that can expand to fit longer substrates. J. Mol. Biol. 365, 135–145 (2007).
pubmed: 17059829 doi: 10.1016/j.jmb.2006.09.049
Tan, Z., Black, W., Yoon, J. M., Shanks, J. V. & Jarboe, L. R. Improving Escherichia coli membrane integrity and fatty acid production by expression tuning of FadL and OmpF. Microb. Cell Factories 16, 1–15 2017
doi: 10.1186/s12934-017-0650-8
Shokri, A. & Larsson, G. Characterisation of the Escherichia coli membrane structure and function during fedbatch cultivation. Microb. Cell Factories 3, 9 (2004).
doi: 10.1186/1475-2859-3-9
Morris, T. W., Reed, K. E. & Cronan, J. E. Lipoic acid metabolism in Escherichia coli: the lplA and lipB genes define redundant pathways for ligation of lipoyl groups to apoprotein. J. Bacteriol. 177, 1–10 (1995).
pubmed: 8002607 pmcid: 176549 doi: 10.1128/jb.177.1.1-10.1995
Majerus, P. W., Alberts, A. W. & Vagelos, P. R. Acyl Carrier Protein, IV. The identification of 4′-phosphopantetheine as the prosthetic group of the acyl carrier protein. Proc. Natl Acad. Sci. USA 53, 410–417 (1965).
pubmed: 14294075 doi: 10.1073/pnas.53.2.410 pmcid: 219528
Zornetzer, G. A., Fox, B. G. & Markley, J. L. Solution structures of spinach acyl carrier protein with decanoate and stearate. Biochemistry 45, 5217–5227 (2006).
pubmed: 16618110 doi: 10.1021/bi052062d
Cronan, J. E. The chain-flipping mechanism of ACP (acyl carrier protein)-dependent enzymes appears universal. Biochem. J. 460, 157–163 (2014).
pubmed: 24825445 doi: 10.1042/BJ20140239
Kim, Y., Kovriguine, E. & Eletr, Z. NMR studies of Escherichia coli acyl carrier protein: dynamic and structural differences of the apo- and holo-forms. Biochem. Biophys. Res. Commun. 17, 776–783 (2006).
doi: 10.1016/j.bbrc.2006.01.025
Andrec, M., Hill, R. B. & Prestegard, J. H. Amide exchange rates in Escherichia coli acyl carrier protein: correlation with protein structure and dynamics. Protein Sci. 4, 983–993 (1995).
pubmed: 7663354 pmcid: 2143121 doi: 10.1002/pro.5560040518
Colizzi, F., Masetti, M., Recanatini, M. & Cavalli, A. Atomic-level characterization of the chain-flipping mechanism in fatty-acids biosynthesis. J. Phys. Chem. Lett. 7, 2899–2904 (2016).
pubmed: 27409360 doi: 10.1021/acs.jpclett.6b01230
Cronan, J. J. & Rock, C. O. Biosynthesis of membrane lipids. EcoSal Plus 3, 3.6.4 (2008).
Liu, T., Vora, H. & Khosla, C. Quantitative analysis and engineering of fatty acid biosynthesis in E. coli. Metab. Eng. 12, 378–386 (2010).
pubmed: 20184964 doi: 10.1016/j.ymben.2010.02.003
Worthington, A. S., Rivera, H., Torpey, J. W., Alexander, M. D. & Burkart, M. D. Mechanism-based protein cross-linking probes to investigate carrier protein-mediated biosynthesis. ACS Chem. Biol. 1, 687–691 (2006).
pubmed: 17184132 doi: 10.1021/cb6003965
Finzel, K. et al. Probing the substrate specificity and protein-protein interactions of the E. coli fatty acid dehydratase, FabA. Chem. Biol. 22, 1453–1460 (2015).
pubmed: 26526101 pmcid: 4656083 doi: 10.1016/j.chembiol.2015.09.009
Miyanaga, A. et al. Structural basis of protein–protein interactions between a trans-acting acyltransferase and acyl carrier protein in polyketide disorazole biosynthesis. J. Am. Chem. Soc. 140, 7970–7978 (2018).
pubmed: 29870659 doi: 10.1021/jacs.8b04162
Miyanaga, A., Iwasawa, S., Shinohara, Y., Kudo, F. & Eguchi, T. Structure-based analysis of the molecular interactions between acyltransferase and acyl carrier protein in vicenistatin biosynthesis. Proc. Natl Acad. Sci. USA 113, 1802–1807 (2016).
pubmed: 26831085 doi: 10.1073/pnas.1520042113 pmcid: 4763789
Weissman, K. J., Hong, H., Popovic, B. & Meersman, F. Evidence for a protein-protein interaction motif on an acyl carrier protein domain from a modular polyketide synthase. Chem. Biol. 13, 625–636 (2006).
pubmed: 16793520 doi: 10.1016/j.chembiol.2006.04.010
Beld, J., Cang, H. & Burkart, M. D. Visualizing the chain-flipping mechanism in fatty acid biosynthesis. Angew. Chem. Int. Ed. Engl. 53, 14456–14461 (2014).
pubmed: 25354391 pmcid: 4425425 doi: 10.1002/anie.201408576
Sarria, S., Bartholow, T. G., Verga, A., Burkart, M. D. & Peralta-Yahya, P. Matching protein interfaces for improved medium-chain fatty acid production. ACS Synth. Biol. 7, 1179–1187 (2018).
pubmed: 29722970 pmcid: 6296256 doi: 10.1021/acssynbio.7b00334
Sztain, T. et al. Modifying the thioester linkage affects the structure of the acyl carrier protein. Angew. Chem. Int. Ed. 58, 10888–10892 (2019).
doi: 10.1002/anie.201903815
Farmer, R., Thomas, C. M. & Winn, P. J. Structure, function and dynamics in acyl carrier proteins. PLoS ONE 14, e0219435 (2019).
pubmed: 31291335 pmcid: 6619796 doi: 10.1371/journal.pone.0219435
Lohman, J. R. et al. The crystal structure of BlmI as a model for nonribosomal peptide synthetase peptidyl carrier proteins. Proteins 82, 1210–1218 (2014).
pubmed: 25050442 pmcid: 4109001 doi: 10.1002/prot.24485
Rajagopal, P., Waygood, E. B., Reizer, J., Saier, M. H. & Klevit, R. E. Demonstration of protein-protein interaction specificity by NMR chemical shift mapping. Protein Sci. Publ. Protein Soc. 6, 2624–2627 (1997).
doi: 10.1002/pro.5560061214
Zuiderweg, E. R. P. Mapping protein−protein interactions in solution by NMR spectroscopy. Biochemistry 41, 1–7 (2002).
pubmed: 11771996 doi: 10.1021/bi011870b
Pellecchia, M. et al. Structural insights into substrate binding by the molecular chaperone DnaK. Nat. Struct. Biol. 7, 298 (2000).
pubmed: 10742174 doi: 10.1038/74062
Williamson, M. P. Using chemical shift perturbation to characterise ligand binding. Prog. Nucl. Magn. Reson. Spectrosc. 73, 1–16 (2013).
pubmed: 23962882 doi: 10.1016/j.pnmrs.2013.02.001
Waudby, C. A. et al. Two-Dimensional NMR Lineshape Analysis. Scientific Reports, 6, 1–8 (2016)
doi: 10.1038/srep24826
Milligan, J. C. et al. Molecular basis for interactions between an acyl carrier protein and a ketosynthase. Nat. Chem. Biol. 15, 669 (2019).
pubmed: 31209348 pmcid: 7323458 doi: 10.1038/s41589-019-0301-y
Nguyen, C. et al. Trapping the dynamic acyl carrier protein in fatty acid biosynthesis. Nature 505, 427–431 (2014).
pubmed: 24362570 doi: 10.1038/nature12810
Mj, J. et al. Manipulating protein-protein interactions in nonribosomal peptide synthetase type II peptidyl carrier proteins. Biochemistry. https://doi.org/10.1021/acs.biochem.7b00884 (2017).
Abagyan, R. & Totrov, M. Biased probability Monte Carlo conformational searches and electrostatic calculations for peptides and proteins. J. Mol. Biol. 235, 983–1002 (1994).
pubmed: 8289329 doi: 10.1006/jmbi.1994.1052
Abagyan, R., Totrov, M. & Kuznetsov, D. ICM—a new method for protein modeling and design: applications to docking and structure prediction from the distorted native conformation. J. Comput. Chem. 15, 488–506 (1994).
doi: 10.1002/jcc.540150503
Mindrebo, J. T. et al. Gating mechanism of elongating β-ketoacyl-ACP synthases. Nat. Commun. 11, 1727 (2020).
pubmed: 32265440 pmcid: 7138838 doi: 10.1038/s41467-020-15455-x
Platensimycin is a selective FabF inhibitor with potent antibiotic properties. Nature 441, 358–361 (2006).
Zhang, Y. M. et al. Identification and analysis of the acyl carrier protein (ACP) docking site on β-ketoacyl-ACP synthase III. J. Biol. Chem. 276, 8231–8238 (2001).
Leesong, M., Henderson, B. S., Gillig, J. R., Schwab, J. M. & Smith, J. L. Structure of a dehydratase-isomerase from the bacterial pathway for biosynthesis of unsaturated fatty acids: two catalytic activities in one active site. Structure 4, 253–264 (1996).
pubmed: 8805534 doi: 10.1016/S0969-2126(96)00030-5
Kozakov, D. et al. The ClusPro web server for protein–protein docking. Nat. Protoc. 12, 255–278 (2017).
pubmed: 28079879 pmcid: 5540229 doi: 10.1038/nprot.2016.169
Vajda, S. et al. New additions to the ClusPro server motivated by CAPRI. Proteins 85, 435–444 (2017).
pubmed: 27936493 pmcid: 5313348 doi: 10.1002/prot.25219
Kozakov, D. et al. How good is automated protein docking? Proteins 81, 2159–2166 (2013).
pubmed: 23996272 pmcid: 3934018 doi: 10.1002/prot.24403
van Zundert, G. C. P. et al. The HADDOCK2.2 Web Server: user-friendly integrative modeling of biomolecular complexes. J. Mol. Biol. 428, 720–725 (2016).
pubmed: 26410586 doi: 10.1016/j.jmb.2015.09.014
Dominguez, C. Boelens, R. & Bonvin, A. M. J. J. HADDOCK: a protein−protein docking approach based on biochemical or biophysical information. J. Am. Chem. Soc. 125, 1731–1737 (2003).
Lyskov, S. et al. Serverification of molecular modeling applications: The Rosetta Online Server That Includes Everyone (ROSIE). PLoS ONE 8, e63906 (2013).
pubmed: 23717507 pmcid: 3661552 doi: 10.1371/journal.pone.0063906
Lyskov, S. & Gray, J. J. The RosettaDock server for local protein–protein docking. Nucleic Acids Res. 36, W233–W238 (2008).
pubmed: 18442991 pmcid: 2447798 doi: 10.1093/nar/gkn216
Joshi, V. C. & Wakil, S. J. Studies on the mechanism of fatty acid synthesis: XXVI. Purification and properties of malonyl-coenzyme A—acyl carrier protein transacylase of Escherichia coli. Arch. Biochem. Biophys. 143, 493–505 (1971).
pubmed: 4934182 doi: 10.1016/0003-9861(71)90234-7
Rafi, S. et al. Structure of acyl carrier protein bound to FabI, the FASII enoyl reductase from Escherichia Coli. J. Biol. Chem. 281, 39285–39293 (2006).
pubmed: 17012233 doi: 10.1074/jbc.M608758200
Garwin, J. L., Klages, A. L. & Cronan, J. E. Structural, enzymatic, and genetic studies of beta-ketoacyl-acyl carrier protein synthases I and II of Escherichia coli. J. Biol. Chem. 255, 11949–11956 (1980).
pubmed: 7002930 doi: 10.1016/S0021-9258(19)70226-9
Yu, X., Liu, T., Zhu, F. & Khosla, C. In vitro reconstitution and steady-state analysis of the fatty acid synthase from Escherichia coli. Proc. Natl Acad. Sci. USA 108, 18643–18648 (2011).
pubmed: 22042840 doi: 10.1073/pnas.1110852108 pmcid: 3219124
Toomey, R. E. & Wakil, S. J. Studies on the mechanism of fatty acid synthesis XV. Preparation and general properties of β-ketoacyl acyl carrier protein reductase from Escherichia coli. Biochim. Biophys. Acta 116, 189–197 (1966).
pubmed: 4381013 doi: 10.1016/0005-2760(66)90001-4
Charov, K. & Burkart, M. D. A single tool to monitor multiple protein–protein interactions of the Escherichia coli acyl carrier protein. ACS Infect. Dis. 5, 1518–1523 (2019).
pubmed: 31317739 pmcid: 7607843 doi: 10.1021/acsinfecdis.9b00150
Zhang, Y.-M., Wu, B., Zheng, J. & Rock, C. O. Key residues responsible for acyl carrier protein and β-ketoacyl-acyl carrier protein reductase (FabG) interaction. J. Biol. Chem. 278, 52935–52943 (2003).
pubmed: 14527946 doi: 10.1074/jbc.M309874200
Ac, P., Ym, Z., Co, R. & Sw, W. Cofactor-induced conformational rearrangements establish a catalytically competent active site and a proton relay conduit in FabG. Structure https://doi.org/10.1016/j.str.2004.02.008 (2004).
Heath, R. J. & Rock, C. O. Enoyl-acyl carrier protein reductase (fabI) plays a determinant role in completing cycles of fatty acid elongation in Escherichia coli. J. Biol. Chem. 270, 26538–26542 (1995).
pubmed: 7592873 doi: 10.1074/jbc.270.44.26538
Cho, H. & Cronan, J. E. ‘Protease I’ of Escherichia coli functions as a thioesterase in vivo. J. Bacteriol. 176, 1793–1795 (1994).
pubmed: 8132479 pmcid: 205273 doi: 10.1128/jb.176.6.1793-1795.1994
Shin, K. S., Kim, S. & Lee, S. K. Improvement of free fatty acid production using a mutant acyl-CoA thioesterase I with high specific activity in Escherichia coli. Biotechnol. Biofuels 9, 208 (2016).
pubmed: 27761152 pmcid: 5053343 doi: 10.1186/s13068-016-0622-y
Delaglio, F., Grzesiek, S., Vuister, G. W., Zhu, G. & Pfeifer, J. A Bax NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J. Biomolecular NMR 6, 277–293 (1995).
Lee, W., Tonelli, M. & Markley, J. L. NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy. Bioinformatics 31, 1325–1327 (2015).
pubmed: 25505092 doi: 10.1093/bioinformatics/btu830
Hunter, J. D. Matplotlib: a 2D graphics environment. Comput. Sci. Eng. 9, 90–95 (2007).
doi: 10.1109/MCSE.2007.55

Auteurs

Thomas G Bartholow (TG)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.

Terra Sztain (T)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.

Ashay Patel (A)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.

D John Lee (DJ)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA, USA.

Megan A Young (MA)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
ORCID: 0000-0002-3330-3580

Ruben Abagyan (R)

Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, USA.

Michael D Burkart (MD)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA. mburkart@ucsd.edu.
ORCID: 0000-0002-4472-2254

Articles similaires

Decoding the genomic terrain: functional insights into 14 chemosensory proteins in whitefly Bemisia tabaci Asia II-1.

M N Rudra Gouda, Sabtharishi Subramanian
1.00
Animals Hemiptera Insect Proteins Phylogeny Insecticides

Membrane potential stimulates ADP import and ATP export by the mitochondrial ADP/ATP carrier due to its positively charged binding site.

Vasiliki Mavridou, Martin S King, Andre Bazzone et al.
1.00
Adenosine Triphosphate Adenosine Diphosphate Mitochondrial ADP, ATP Translocases Binding Sites Mitochondria

Conservation of the cooling agent binding pocket within the TRPM subfamily.

Kate Huffer, Matthew C S Denley, Elisabeth V Oskoui et al.
1.00
TRPM Cation Channels Animals Binding Sites Mice Pyrimidinones

Drug repurposing against fucosyltransferase-2 via docking, STD-NMR, and molecular dynamic simulation studies.

Muhammad Atif, Humaira Zafar, Atia-Tul- Wahab et al.
1.00
Fucosyltransferases Drug Repositioning Molecular Docking Simulation Molecular Dynamics Simulation Humans

Classifications MeSH

questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Acyltransferases › 3-Oxoacyl (acyl carrier protein) synthase › Elucidation of transient protein-protein...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Acyltransferases › Acetyltransferases › Elucidation of transient protein-protein...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de transport › Protéine ACP › Elucidation of transient protein-protein...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Oxidoreductases › Alcohol oxidoreductases › Elucidation of transient protein-protein...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Sites de fixation › Elucidation of transient protein-protein...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Oxidoreductases › Oxidoreductases acting on CH-CH group donors › Enoyl-(acyl-carrier protein) reductase (NADH) › Elucidation of transient protein-protein...
questionsmedicales.fr › Bactéries › Proteobacteria › Gammaproteobacteria › Enterobacteriaceae › Escherichia › Escherichia coli › Elucidation of transient protein-protein...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Protéines Escherichia coli › Elucidation of transient protein-protein...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Acyltransferases › Acyl-carrier protein S-malonyltransferase › Fatty acid synthases › Fatty acid synthase type II › Elucidation of transient protein-protein...
questionsmedicales.fr › Lipides › Acides gras › Elucidation of transient protein-protein...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Esterases › Carboxylic ester hydrolases › Lysophospholipase › Elucidation of transient protein-protein...
questionsmedicales.fr › Sciences de l'information › Méthodologies informatiques › Simulation numérique › Simulation de docking moléculaire › Elucidation of transient protein-protein...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines végétales › Protéines périplasmiques › Elucidation of transient protein-protein...
questionsmedicales.fr › Métabolisme › Liaison aux protéines › Elucidation of transient protein-protein...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Conformation moléculaire › Conformation des protéines › Éléments structuraux des protéines › Motifs et domaines d'intéraction protéique › Elucidation of transient protein-protein...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Analyse spectrale › Spectroscopie par résonance magnétique › Spectroscopie par résonance magnétique du proton › Elucidation of transient protein-protein...