Strains, plasmids, and polymerase chain reaction (PCR) primers used
in this study are summarized in Tables S1–S3, respectively,
of the
Supporting Information.
Escherichia coli XL1 Blue MR was used for common subcloning
and plasmid preparation.
33 E. coli BL21(DE3) was used for protein overproduction. Oligonucleotide primer
synthesis and DNA sequencing were performed by the University of Wisconsin—Madison
Biotechnology Center. The
Expand High Fidelity PCR System (Roche)
was used for PCR amplification. Commerical kits (Promega) were used
for gel extraction and plasmid preparation. All restriction endonucleases
and T4 DNA ligase were purchased from NEB and reactions conducted
according to the manufacturer’s protocols. Cu(II)-ZBM and metal-free
TLM A were isolated from
S. flavoviridis SB9001 and
Sa. hindustanus E465-94 ATCC31154, respectively, as previously
reported.
16 (link),30 (link) Cu(II)-BLM B2 (Nippon Kayaku, Tokyo, Japan),
Cu(II)-PLM D1 (Cayla, Toulouse, France),
metal-free BLM B2 (VWR International),
[
acetyl-1-
14C]acetyl-CoA (Moravek Biochemicals),
and
EcoLume scintillation cocktail (MP Biomedicals) were purchased
from commercial sources. Other common biochemicals and chemicals were
purchased from standard commercial sources and used directly.
Coughlin J.M., Rudolf J.D., Wendt-Pienkowski E., Wang L., Unsin C., Galm U., Yang D., Tao M, & Shen B. (2014). BlmB and TlmB Provide Resistance to the Bleomycin Family of Antitumor Antibiotics by N-Acetylating Metal-Free Bleomycin, Tallysomycin, Phleomycin, and Zorbamycin. Biochemistry, 53(44), 6901-6909.
