O1-6

SynergyUltra opens a new era with digits of different
performances and capabilities in protein crystallography;
4s-4m-40m
Takashi Matsumoto 1

Kazuaki Aburaya 1

Takashi Sato 1

t-matumo@rigaku.co.jp

aburaya@rigaku.co.jp

satow@rigaku.ac.jp

1

Rigaku Corporation, 3-9-12 Matsubara-cho, Akishima, Tokyo 196-8666, Japan

Keywords: SynergyUltra, 4 seconds whole measurement, 4s-4m-40m
The accurate protein structures and their complex targets compounds are necessary and criticalfor designing and developing effective medicines and potent substances. To date, massive numbersof these structures have been determined by single -crystal X-ray diffraction (SC-XRD) analysis.
Other new technologies like single-particle analysis using a Cryo-electron microscope or, more
recently, utilization of three-dimensional electron diffraction (3D-ED) are emerging to overcome
the difficulty of crystallization and hence to be thought the enhancement in the speed.
The geometry of the atoms defines interactions among t he molecules, and some of those arehighly sensitive to the directions and/or distances; thus, the accurate and precise structur alinformation of the molecule is an indispensable firm base for successful drug design processes. In
this context, SC-XRD is still the best approach if it could be performed as its necessity.
Meanwhile, it has still been widely believed that SC-XRD requires considerable time and effort tocrystalize the target molecules and other efforts to collect the datasets and phasing.
This presentation introduces our next-generation instrument that opens a new era of protein
SC-XRD with laboratory instruments formats. Ultra-fast and accurate measurements of protein
crystals using our latest system SynergyUltra will be presented. The following numbers
characterize the power of SynergyUltra.
-4 seconds, the whole data collection for the molecular replacement phasing:The 4 seconds in total data collection fulfilling the 1.8Å resolution range. The obtained
results were accurate enough for molecular replacement phasing employing Alpha Fold 2
deduced structure as a searching model. As a result, detailed electron densities wereobtained, in which disorder of the sidechain can be modeled and refined for theirpopulation. Utilizing the SynergyFlow robotics system enables to obtain the unique crystal
structures every several minutes that enable curtain bombing (carpet bombing) structure
analyses of mutant proteins and/or its s complexes with various candidates.

-4 minutes, the whole data collection good for SAD phasing:The 4 minutes in total data collection fulfill ing the 1.75Å resolution range. Thismeasurement allows obtaining highly even and redundant data sets that are accurate enoughfor successful S-SAD phasing. 97% of residues were automatically modeled and refined toyield the detailed electron densities. 4 minutes measurements would be useful unless above
4 seconds MR experiments did not work well.

-40 minutes, the whole data collection for ultra -high resolution data collection.
The 40 minutes in total data collection fulfillin g the 1.40Å resolution range. Thismeasurement allows obtaining further precise and highly accurate electron density thatshows the spherical shapes for each atom similar to those obtained in small -molecule
crystallography in that atom types can be identified by their density.