000170128 001__ 170128
000170128 005__ 20260318155255.0
000170128 0247_ $$2doi$$a10.1038/s42004-026-01908-9
000170128 0248_ $$2sideral$$a148656
000170128 037__ $$aART-2026-148656
000170128 041__ $$aeng
000170128 100__ $$aDoppler, Diandra
000170128 245__ $$aMinimized sample consumption for time-resolved serial crystallography applied to the redox cycle of human NQO1
000170128 260__ $$c2026
000170128 5060_ $$aAccess copy available to the general public$$fUnrestricted
000170128 5203_ $$aSample consumption for serial femtosecond crystallography with X-ray free electron lasers remains a major limitation preventing broader use in macromolecular crystallography. This drawback is exacerbated in time-resolved (TR) experiments, where the amount of sample required per reaction time point is multiplied by the number of time points investigated. To reduce this limitation, we demonstrate a segmented droplet generation strategy coupled to a mix-and-inject approach for TR studies at the European XFEL. The injector produces synchronized droplet trains that enable stable and reproducible injection of protein crystal slurries at significantly reduced flow rates. Using the human flavoenzyme NAD(P)H:quinone oxidoreductase 1 (NQO1) as a test system, we collected diffraction data after mixing with NADH at 0.3 s and 1.2 s delays. The segmented injection approach achieved up to 97% reduction in sample consumption compared with continuous-flow injection while maintaining data quality suitable for TR crystallography. Reproducible electron density features consistent with low-occupancy NADH binding illustrate both the feasibility and the current limits of studying dynamic redox enzymes using this approach. This work establishes segmented droplet generation as a sample-efficient and XFEL-compatible method for future time-resolved serial crystallography experiments.
000170128 536__ $$9info:eu-repo/grantAgreement/ES/DGA/E35-23R$$9info:eu-repo/grantAgreement/ES/MICINN/CNS2022-135713$$9info:eu-repo/grantAgreement/ES/MICINN/PID2022-136369NB-I00$$9info:eu-repo/grantAgreement/ES/MICINN/PID2023-151100NB-I00
000170128 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttps://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
000170128 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000170128 700__ $$aGrieco, Alice
000170128 700__ $$aKoh, Domin
000170128 700__ $$aManna, Abhik
000170128 700__ $$aAnsari, Adil
000170128 700__ $$aAlvarez, Roberto
000170128 700__ $$aKarpos, Konstantinos
000170128 700__ $$aLe, Hung
000170128 700__ $$aSonker, Mukul
000170128 700__ $$aKetawala, Gihan K.
000170128 700__ $$aMahmud, Samira
000170128 700__ $$aQuereda-Moraleda, Isabel
000170128 700__ $$aSen, Sayantee
000170128 700__ $$aPey, Angel L.
000170128 700__ $$aLetrun, Romain
000170128 700__ $$aDörner, Katerina
000170128 700__ $$aKoliyadu, Jayanath C. P.
000170128 700__ $$ade Wijn, Raphael
000170128 700__ $$aBielecki, Johan
000170128 700__ $$aHan, Huijong
000170128 700__ $$aKim, Chan
000170128 700__ $$aKoua, Faisal H. M.
000170128 700__ $$aRound, Adam
000170128 700__ $$aSarma, Abhisakh
000170128 700__ $$aSato, Tokushi
000170128 700__ $$aSchmidt, Christina
000170128 700__ $$aVakili, Mohammad
000170128 700__ $$aZabelskii, Dmitrii
000170128 700__ $$aBean, Richard
000170128 700__ $$aMancuso, Adrian P.
000170128 700__ $$aSchulz, Joachim
000170128 700__ $$aFromme, Raimund
000170128 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, Milagros$$uUniversidad de Zaragoza
000170128 700__ $$aGrant, Thomas D.
000170128 700__ $$aFromme, Petra
000170128 700__ $$aKirian, Richard A.
000170128 700__ $$aBotha, Sabine
000170128 700__ $$aManuel Martin-Garcia, Jose
000170128 700__ $$aRos, Alexandra
000170128 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000170128 773__ $$g9, 1 (2026), 14 pp.$$tCommunications Chemistry$$x2399-3669
000170128 8564_ $$s3341953$$uhttps://zaguan.unizar.es/record/170128/files/texto_completo.pdf$$yVersión publicada
000170128 8564_ $$s2774976$$uhttps://zaguan.unizar.es/record/170128/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000170128 909CO $$ooai:zaguan.unizar.es:170128$$particulos$$pdriver
000170128 951__ $$a2026-03-18-13:52:53
000170128 980__ $$aARTICLE