REL 2023-08-12 2024-01-15 2024-01-15 0000-0001-6653-6682 Mark A Herzik Jr Department of Chemistry and Biochemistry, University of California, San Diego 9500 Gilman Drive La Jolla California United States 92093 0000-0001-6653-6682 Mark A Herzik Jr Department of Chemistry and Biochemistry, University of California, San Diego 9500 Gilman Drive La Jolla California United States 92093 0000-0002-4733-6500 Faik Akif Tezcan Department of Chemistry and Biochemistry, University of California, San Diego 9500 Gilman Drive La Jolla California United States 92093 Rutledge HL Cook BD Nguyen HPM Tezcan FA Herzik Jr MA National Institutes of Health (NIH) R01-GM099813 United States National Institutes of Health (NIH) R35-GM138206 United States National Institutes of Health (NIH) T32-GM008326 United States National Aeronautic Space Administration (NASA, United States) ENIGMA United States Kinship Foundation Searle Scholars Program United States 5.8 10.6019/EMPIAR-11795 EMDB molecule EMD-26764 Rutledge HL Cook BD Nguyen HPM Herzik Jr MA Tezcan FA Science (New York, N.Y.) Science 6608 377 865 869 2022 English 10.1126/science.abq7641 35901182

CryoEM movies of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions in the presence of the inhibitor of berrylium fluoride (BeFx). Data were collected on a G3 Titan Krios (300 kV) at a nominal magnification of 165,000x (0.815 Å/pixel) in EFTEM mode on a Gatan K2 detector post a Quantum energy filter (20 eV slit) and a cumulative electron exposure of ~65 electrons/Å2 (50 frames) recorded in counting mode. Data were collected automatically using EPU with aberration free image shift using a defocus range of -0.5 – -2.5 μm. Four separate data sets were collected using 0˚, 15˚ or 25˚ specimen tilt.

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx. /data/set1_0tilt/movies micrographs - multiframe MRC MRC 2198 50 UNSIGNED BYTE 3838 0.815 3710 0.815

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames).

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx. /data/set1_0tilt/micrographs micrographs - single frame MRC MRC 2198 1 32 BIT FLOAT 3838 0.815 3710 0.815

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames).

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx /data/set2_0tilt/movies micrographs - multiframe MRC MRC 1718 50 UNSIGNED BYTE 3838 0.815 3710 0.815

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames).

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx /data/set2_0tilt/micrographs micrographs - single frame MRC MRC 1718 1 32 BIT FLOAT 3838 0.815 3710 0.815

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames).

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx at 15 degree tilt /data/set3_15tilt/movies micrographs - multiframe MRC MRC 2211 50 UNSIGNED BYTE 3838 0.815 3710 0.815

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames). Stage was tilted at 15 degrees.

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx at 15 degree tilt /data/set3_15tilt/micrographs micrographs - single frame MRC MRC 2211 1 32 BIT FLOAT 3838 0.815 3710 0.815

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames). Stage was tilted at 15 degrees.

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx at 25 degree tilt /data/set4_25tilt/movies micrographs - multiframe MRC MRC 2103 50 UNSIGNED BYTE 3838 0.815 3710 0.815

Unaligned multi-frame micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames). Stage was tilted at 25 degrees.

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx at 25 degree tilt /data/set4_25tilt/micrographs micrographs - single frame MRC MRC 2103 1 32 BIT FLOAT 3838 0.815 3710 0.815

Aligned micrographs of nitrogenase (MoFeP + FeP) from Azotobacter vinelandii under catalytic turnover conditions with inhibitor BeFx collected on a Titan Krios G3 (Thermo Fisher Scientific) operating at 300 keV equipped with a Gatan BioContinuum energy filter. Images were collected at a magnification of 165,000x in EF-TEM mode (0.815 Å calibrated pixel size) on a Gatan K2 detector using a 20-eV slit width and a cumulative electron exposure of ~65 electrons/Å2 (50 frames). Stage was tilted at 25 degrees.