Ringdahl U, Svensson HG, Kotarsky H, Gustafsson M, Weineisen M, Sjobring U. the Innovative Commons Attribution 4.0 International permit. FIG?S3. Kinetic evaluation of IgA binding to immobilized M28 and M1 and C4BP binding to M28 suited VAV1 to different versions. (A) IgA binding to immobilized M28 fitted to a 1-to-1 model (I) and a heterogeneous ligand model CZC-25146 hydrochloride (II). (B) IgA binding to immobilized M1 fitted to a 1-to-1 model (I) and a heterogeneous ligand model (II). (C) C4BP binding to immobilized M28 fitted to a 1-to-1 model (I) and a heterogeneous ligand model (II). (D) Chi-squared values for 1-to-1 binding and heterogeneous ligand models for M28-IgA, M1-IgA, and M28-C4BP interactions. Download FIG?S3, PDF file, 0.6 MB. Copyright ? 2021 Chowdhury et al. This content is distributed under the terms CZC-25146 hydrochloride of the Creative Commons Attribution 4.0 International license. FIG?S4. Individual spectra for identified cross-linked peptides. (A and B) Spectra for the M2-C4BP interface; (C and D) spectra for the M28-IgA interface overlapping the identified C4BP interaction interface and the previously identified M22-based IgA-binding SAP; (E to L) spectra for the novel M28-IgA interface. The cross-linked peptides are indicated at the top of each spectrum. The fragments indicated in red and blue are from the individual parent peptides, whereas the fragments in green arise from a fragmented cross-linked peptide. The intensity for each spectrum is shown on the axis, and the ratio is shown on the axis. Download FIG?S4, PDF file, 0.4 MB. Copyright ? 2021 Chowdhury et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S1. M protein sequences and UniProt identifiers. The protein sequences of different M proteins expressed recombinantly along with their UniProt identifiers are presented. The initiating methionine residue is marked in red, and the colored sequence represents the tag incorporated into the protein. Download Table?S1, DOCX file, 0.1 MB. Copyright ? 2021 Chowdhury et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S2. List of peptides from M28 cross-linked to peptides from either C4BP or IgA Fc. The cross-linked lysine residues (K) are in boldface type. Download Table?S2, DOCX file, 0.09 MB. Copyright ? 2021 Chowdhury et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S5. IgA binding to GFP, M28, and M89. (A) Multiple-sequence alignment of M28 and M89. Purple denotes the SAP sequence, which is present only in M28, and red denotes the novel IgA-binding site, which is conserved in both M28 and M89. (B) DIA of AP-MS for IgA from human plasma. Intensity is represented as a ratio of the sum of the intensity of the top three peptides of IgA to the sum of all the peptides identified during AP-MS of GFP, M28, and M89 with human plasma. M89 binds to IgA but with a lower affinity than M28. (C) DDA of AP-MS for commercial IgA. Intensity is represented as a ratio of the sum of the intensity of the top three peptides of IgA to the sum of all the peptides identified during AP-MS of commercial IgA with GFP, M28, and M89. M89 seems to bind IgA with a lower affinity than M28. Download FIG?S5, EPS file, 1.8 MB. Copyright ? 2021 Chowdhury et al. CZC-25146 hydrochloride This CZC-25146 hydrochloride content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S1. Multiple-sequence alignment of mature M proteins. Shown is a multiple-sequence alignment of the amino acid sequences of M1, M3, M28, M49, and M89 proteins recombinantly expressed and used for AP-MS experiments. The purple-highlighted region represents the SAP sequence on M28. The red region represents the novel identified IgA Fc-binding site on M28. Download FIG?S1, EPS file, 2.4 MB. Copyright ? 2021 Chowdhury et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. Reviewer comments reviewer-comments.pdf (605K) GUID:?DEC07F03-8ECF-454B-890B-776E5E71AE66 ABSTRACT is known to cause both mucosal and systemic infections in humans. In this study, we used a combination of quantitative and structural mass spectrometry techniques to determine the composition and structure of the interaction network formed between human plasma proteins and the surfaces of different serotypes. Quantitative network analysis revealed that forms serotype-specific interaction.