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Actinobacillus seminis GroEL-homologous protein agglutinates sheep erythrocytes

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Abstract

Actinobacillus seminis, a commensal of ovine and caprine reproductive organs, is able to induce epididymitis in the small ruminants that it infects. In this work, we characterised two protein bands of approximately 150 kDa and 65 kDa. These proteins cross-reacted with a polyclonal serum against Gallibacterium anatis hemagglutinin and with a polyclonal serum from sheep with epididymitis, indicating that the proteins are expressed in vivo; the two proteins also interacted with biotin-labeled sheep fibrinogen and fibronectin, suggesting that they may function as adhesins. The participation of these proteins as adhesins was confirmed by a cultured human bladder cell-A. seminis adhesion assay and adherence inhibition by preincubation of A. seminis with polyclonal antiserum to the 150 kDa protein. Both proteins presented sequence identity with an A. seminis GroEL protein by mass spectrometry analysis and agglutinated glutaraldehyde-fixed sheep red blood cells. Immunogold labeling was observed by transmission electron microscopy on bacterial cells that were negatively stained, and a peroxidase reaction was detected in A. seminis biofilms, when an anti-A. seminis 150 kDa protein serum was used, indicating the presence of this protein on the surface of A. seminis and in biofilms. The A. seminis GroEL-homologue is a multifunctional protein that likely acts as a hemagglutinin.

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Abbreviations

EF-Tu:

Elongation factor-Tu

PGM:

Phosphoglycerate mutase

HA:

Hemagglutinin

OMP:

Outer membrane protein

TEM:

Transmission electron microscopy

TS:

Tryptic soy

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Acknowledgements

This study was funded by grants from DGAPA-UNAM PAPIIT IN215616 and IN219919. FMG is a doctoral student supported by a scholarship from CONACYT (283503).

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Author notes

  1. Sergio Vaca: Died February, 2019.

Authors and Affiliations

  1. Carrera de Biología, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Av. de los Barrios # 1, Los Reyes Iztacala, 54090, Tlalnepantla, Estado de México, Mexico

    Juan Fernando Montes-García, Willy Angel Delgado-Tapia, Sergio Vaca & Erasmo Negrete-Abascal

  2. Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, BUAP, Apdo. Postal 1622, 72560, Puebla, Mexico

    Candelario Vazquez-Cruz

  3. Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México “Federico Gómez”, Ciudad de México, Mexico

    Ariadnna Cruz-Córdova

Authors
  1. Juan Fernando Montes-García
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  2. Willy Angel Delgado-Tapia
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  3. Candelario Vazquez-Cruz
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  4. Sergio Vaca
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  5. Ariadnna Cruz-Córdova
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  6. Erasmo Negrete-Abascal
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Correspondence to Erasmo Negrete-Abascal.

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Montes-García, J.F., Delgado-Tapia, W.A., Vazquez-Cruz, C. et al. Actinobacillus seminis GroEL-homologous protein agglutinates sheep erythrocytes. Antonie van Leeuwenhoek 112, 1655–1662 (2019). https://doi.org/10.1007/s10482-019-01292-6

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