PMID- 31108423
OWN - NLM
STAT- MEDLINE
DCOM- 20191029
LR  - 20191029
IS  - 1872-9142 (Electronic)
IS  - 0161-5890 (Linking)
VI  - 112
DP  - 2019 Aug
TI  - Structural and immunological characterization of a new 
      nucleotidyltransferase-like antigen from Paracoccidioides brasiliensis.
PG  - 151-162
LID - S0161-5890(19)30012-4 [pii]
LID - 10.1016/j.molimm.2019.04.028 [doi]
AB  - Pb27 antigen is an interesting alternative to immunological diagnosis of 
      Paracoccidioidomycosis (PCM) and has demonstrated to be protective in 
      experimental PCM. Its tertiary structure and possible function remained unknown 
      till now. To study Pb27 at the atomic level, the recombinant protein was 
      expressed in Escherichia coli BL21(DE3), purified, and its three-dimensional 
      structure was solved by X-ray crystallography. Based on this structure, we 
      performed a residue correlation analysis and in silico ligand search assays to 
      address a possible biological function to Pb27. We identified Pb27 as a member of 
      the extensive nucleotidyltransferase superfamily. The protein has an alphabetaalphabetaalphabeta 
      topology with two domains (N- and C-terminal domains) and adopts a monomeric form 
      as its biological unit in solution. Structural comparisons with similar members 
      of the superfamily clearly indicate Pb27 C-terminal domain is singular and may 
      play an important role in its biological function. Bioinformatics analysis 
      suggested that Pb27 might bind to ATP and CTP. This suggestion is corroborated by 
      the fact that a magnesium cation is coordinated by two aspartic acid residues 
      present at the active site (between N- and C-terminal domains), as evidenced by 
      X-ray diffraction data. Besides, NMR assays ((1)H-(15)N HSQC spectra) confirmed 
      the binding of CTP to Pb27, demonstrating for the first time an interaction 
      between a nucleotide and this protein. Moreover, we evaluated the reactivity of 
      sera from patients with Paracoccidioides brasiliensis infection against the 
      recombinant form of Pb27 and showed that it was recognized by sera from infected 
      and treated patients. Predicted B and T cell epitopes were synthesized and 
      further evaluated against sera of PCM patients, providing information of the most 
      reactive peptides in Pb27 primary structure which interact with specific Pb27 
      antibodies.
CI  - Copyright (c) 2019 Elsevier Ltd. All rights reserved.
FAU - Coitinho, Juliana B
AU  - Coitinho JB
AD  - Departamento de Ciencias Fisiologicas, Centro de Ciencias da Saude, Universidade 
      Federal do Espirito Santo, Vitoria, ES 29043-900, Brazil.
FAU - Costa, Mariana A F
AU  - Costa MAF
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - Melo, Eliza M
AU  - Melo EM
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - Morais, Elis A
AU  - Morais EA
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - de Andrade, Lorena G A
AU  - de Andrade LGA
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - da Rocha, Aline M
AU  - da Rocha AM
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - de Magalhaes, Mariana T Q
AU  - de Magalhaes MTQ
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - Favaro, Denize C
AU  - Favaro DC
AD  - Instituto de Quimica, Universidade Estadual de Campinas, Campinas, SP 13083-970, 
      Brazil.
FAU - Bleicher, Lucas
AU  - Bleicher L
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - Pedroso, Enio R P
AU  - Pedroso ERP
AD  - Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, MG 
      31270-901, Brazil.
FAU - Goes, Alfredo M
AU  - Goes AM
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil; 
      Departamento de Patologia Geral, Instituto de Ciencias Biologicas, Universidade 
      Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
FAU - Nagem, Ronaldo A P
AU  - Nagem RAP
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, 
      Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil. 
      Electronic address: nagem@icb.ufmg.br.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190517
PL  - England
TA  - Mol Immunol
JT  - Molecular immunology
JID - 7905289
RN  - 0 (Epitopes, B-Lymphocyte)
RN  - 0 (Epitopes, T-Lymphocyte)
RN  - 0 (Fungal Proteins)
RN  - 0 (Pb27 protein, Paracoccidioides brasiliensis)
RN  - 0 (Recombinant Proteins)
RN  - 65-47-4 (Cytidine Triphosphate)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.7.- (Nucleotidyltransferases)
SB  - IM
MH  - Adenosine Triphosphate/immunology
MH  - Adolescent
MH  - Adult
MH  - Aged
MH  - Amino Acid Sequence
MH  - Cytidine Triphosphate/immunology
MH  - Epitopes, B-Lymphocyte/immunology
MH  - Epitopes, T-Lymphocyte/immunology
MH  - Escherichia coli/immunology
MH  - Female
MH  - Fungal Proteins/*immunology
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Nucleotidyltransferases/*immunology
MH  - Paracoccidioides/*immunology
MH  - Paracoccidioidomycosis/*immunology
MH  - Recombinant Proteins/immunology
MH  - Young Adult
OTO - NOTNLM
OT  - B and T cells epitopes
OT  - CTP binding protein
OT  - Nuclear magnetic resonance
OT  - Paracoccidioidomycosis
OT  - Pb27 nucleotidyltransferase-like antigen
OT  - X-ray diffraction
EDAT- 2019/05/21 06:00
MHDA- 2019/10/30 06:00
CRDT- 2019/05/21 06:00
PHST- 2019/01/10 00:00 [received]
PHST- 2019/04/29 00:00 [revised]
PHST- 2019/04/30 00:00 [accepted]
PHST- 2019/05/21 06:00 [pubmed]
PHST- 2019/10/30 06:00 [medline]
PHST- 2019/05/21 06:00 [entrez]
AID - S0161-5890(19)30012-4 [pii]
AID - 10.1016/j.molimm.2019.04.028 [doi]
PST - ppublish
SO  - Mol Immunol. 2019 Aug;112:151-162. doi: 10.1016/j.molimm.2019.04.028. Epub 2019 
      May 17.