TY - JOUR
T1 - Characterization of noncovalent protein-ligand complexes and associated enzyme intermediates of GlcNAc-6-O-Sulfotransferase by electrospray ionization FT-ICR mass spectrometry
AU - Yu, Yonghao
AU - Kirkup, Colleen E.
AU - Pi, Na
AU - Leary, Julie A.
N1 - Funding Information:
The authors gratefully acknowledge funding for this research by a grant from the National Institute of Health, GM 63581.
PY - 2004/10
Y1 - 2004/10
N2 - In this study, a GlcNAc-6-O-Sulfotransferase, NodST and its complexation with the substrate 3′-phosphoadenosine 5′-phosphosulfate (PAPS) and the inhibitor 3′-phosphoadenosine 5′-phosphate (PAP) were studied using Fourier transform ion cyclotron resonance (FTICR) mass spectrometry. In addition, using isotopically labeled substrate, we have successfully confirmed a sulfated enzyme intermediate, which was predicted by the MS kinetic measurement. It is also shown that information regarding solution binding affinities can be obtained using electrospray ionization (ESI)-FTICR mass spectrometry. The relative binding constants, Kd(PAPS)/K d(PAP), derived from the solution and gas phase were very similar, which suggests that the binding domain of this particular enzyme system, given known structures of other sulfotransferases, may be preserved during the transmission of the complex from solution to the gas phase.
AB - In this study, a GlcNAc-6-O-Sulfotransferase, NodST and its complexation with the substrate 3′-phosphoadenosine 5′-phosphosulfate (PAPS) and the inhibitor 3′-phosphoadenosine 5′-phosphate (PAP) were studied using Fourier transform ion cyclotron resonance (FTICR) mass spectrometry. In addition, using isotopically labeled substrate, we have successfully confirmed a sulfated enzyme intermediate, which was predicted by the MS kinetic measurement. It is also shown that information regarding solution binding affinities can be obtained using electrospray ionization (ESI)-FTICR mass spectrometry. The relative binding constants, Kd(PAPS)/K d(PAP), derived from the solution and gas phase were very similar, which suggests that the binding domain of this particular enzyme system, given known structures of other sulfotransferases, may be preserved during the transmission of the complex from solution to the gas phase.
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U2 - 10.1016/j.jasms.2004.06.002
DO - 10.1016/j.jasms.2004.06.002
M3 - Article
C2 - 15465352
AN - SCOPUS:4744338745
SN - 1044-0305
VL - 15
SP - 1400
EP - 1407
JO - Journal of the American Society for Mass Spectrometry
JF - Journal of the American Society for Mass Spectrometry
IS - 10
ER -