Mass spectra of reactions incubated with WbpB, WbpE, and WbpD.
a, the substrate, UDP-d-GlcNAcA (621.34 g/mol), yields a
mass spectrum in negative ion mode with observed peaks at 620.02
m/z and its natural abundant isotopes at ∼621, ∼622,
and ∼623 m/z. b, after incubation with both WbpB and
WbpE, two additional peaks appear at 618.01 and 619.04 m/z
(marked by an asterisk), corresponding to
UDP-3-keto-d-GlcNAcA (619.33 g/mol) and UDP-d-GlcNAc3NA
(620.36 g/mol), respectively. Neither peak was observed when either WbpB or
WbpE was reacted alone (data not shown). c, enlarged view of
panel b, showing the peaks that correspond to
UDP-3-keto-d-GlcNAcA and UDP-d-GlcNAc3NA. d,
after incubation with WbpB, WbpE, and WbpD, a new peak not present in the
no-enzyme control appears at 661.20 m/z corresponding to
UDP-d-GlcNAc3NAcA (662.40 g/mol). The 661 m/z
peak was not observed when WbpD was absent from the reaction mixture (data not
shown).