Substoichiometric
Ab binding to the S-trimer is sufficient to neutralize
receptor binding. (A–C) MP and CD-MS histograms of ACE2 alone,
revealing the dimeric nature of the utilized ACE2 construct and (D–F)
ACE2 binding to the S-trimer. These results show that ACE2 is largely
dimeric, and only the ACE2-dimer binds to S-trimer, whereby the S-trimer
can accommodate either one or two ACE2. (G–L) MP and CD-MS
histograms of ACE2 binding to the S-trimer following preincubation
with either (G–I) COVA2-15 or (J–L) COVA1-18. The observed
mass shifts of ∼150 kDa (and not 200 kDa) indicate that both
Abs fully prevent ACE2 binding to the S-trimer. Mixing ratios of 4:1
and 4:4:1 (ACE2-dimer:S-trimer and Ab:ACE2-dimer:S-trimer, respectively)
were used for the CD-MS experiments, while 1:1 and 3:1:1 were used
for the MP experiments. Note the similarities between the data presented
in panel G and Figure 2A, and panel J
and Figure 2B.