Carboplatin binding to a model protein in non-NaCl conditions to
eliminate partial conversion to cisplatin, and the use of different criteria
to choose the resolution limit
Hen egg white lysozyme (HEWL) co-crystallisation conditions of carboplatin
without sodium chloride (NaCl) have been utilised to eliminate partial
conversion of carboplatin to cisplatin observed previously. Tetragonal HEWL
crystals were successfully obtained in 65% MPD with 0.1M citric acid buffer at
pH 4.0 including DMSO. The X-ray diffraction data resolution to be used for the
model refinement was reviewed using several topical criteria together. The
CC1/2 criterion implemented in XDS led to data being significant to 2.0{\AA},
compared to the data only being able to be processed to 3.0{\AA} using the
Bruker software package (SAINT). Then using paired protein model refinements
and DPI values based on the FreeR value, the resolution limit was fine tuned to
be 2.3{\AA}. Interestingly this was compared with results from the EVAL
software package which gave a resolution limit of 2.2{\AA} solely using
crossing 2, but 2.8{\AA} based on the Rmerge values (60%). The
structural results showed that carboplatin bound to only the N{\delta} binding
site of His-15 one week after crystal growth, whereas five weeks after crystal
growth, two molecules of carboplatin are bound to the His-15 residue. In
summary several new results have emerged: - firstly non-NaCl conditions showed
a carboplatin molecule bound to His-15 of HEWL; secondly binding of one
molecule of carboplatin was seen after one week of crystal growth and two
molecules were bound after five weeks of crystal growth; and thirdly the use of
several criteria to determine the diffraction resolution limit led to the
successful use of data to higher resolution.Comment: 14 pages; submitted to Acta Cryst D Biological Crystallography
reference number tz504
