Backbone NMR resonance assignments for the C terminal domain of the Streptococcus mutans adhesin P1.
C3 domain
NMR
Quaternary interaction
Streptococcus mutans adhesin P1
Journal
Biomolecular NMR assignments
ISSN: 1874-270X
Titre abrégé: Biomol NMR Assign
Pays: Netherlands
ID NLM: 101472371
Informations de publication
Date de publication:
12 2023
12 2023
Historique:
received:
11
09
2023
accepted:
06
10
2023
medline:
8
11
2023
pubmed:
21
10
2023
entrez:
21
10
2023
Statut:
ppublish
Résumé
Adhesin P1 (aka AgI/II) plays a pivotal role in mediating Streptococcus mutans attachment in the oral cavity, as well as in regulating biofilm development and maturation. P1's naturally occurring truncation product, Antigen II (AgII), adopts both soluble, monomeric and insoluble, amyloidogenic forms within the bacterial life cycle. Monomers are involved in important quaternary interactions that promote cell adhesion and the functional amyloid form promotes detachment of mature biofilms. The heterologous, 51-kD C123 construct comprises most of AgII and was previously characterized by X-ray crystallography. C123 contains three structurally homologous domains, C1, C2, and C3. NMR samples made using the original C123 construct, or its C3 domain, yielded moderately resolved NMR spectra. Using Alphafold, we re-analyzed the P1 sequence to better identify domain boundaries for C123, and in particular the C3 domain. We then generated a more tractable construct for NMR studies of the monomeric form, including quaternary interactions with other proteins. The addition of seven amino acids at the C-terminus greatly improved the spectral dispersion for C3 relative to the prior construct. Here we report the backbone NMR resonance assignments for the new construct and characterize some of its quaternary interactions. These data are in good agreement with the structure predicted by Alphafold, which contains additional β-sheet secondary structure compared to the C3 domain in the C123 crystal structure for a construct lacking the seven C-terminal amino acids. Its quaternary interactions with known protein partners are in good agreement with prior competitive binding assays. This construct can be used for further NMR studies, including protein-protein interaction studies and assessing the impact of environmental conditions on C3 structure and dynamics within C123 as it transitions from monomer to amyloid form.
Identifiants
pubmed: 37864759
doi: 10.1007/s12104-023-10158-y
pii: 10.1007/s12104-023-10158-y
pmc: PMC10695118
mid: NIHMS1943048
doi:
Substances chimiques
Adhesins, Bacterial
0
Amyloid
0
Amino Acids
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
293-299Subventions
Organisme : NIDCR NIH HHS
ID : R01 DE021789
Pays : United States
Organisme : NIH HHS
ID : S10 OD028753
Pays : United States
Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer Nature B.V.
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