Limits...
EEVD motif of heat shock cognate protein 70 contributes to bacterial uptake by trophoblast giant cells.

Watanabe K, Tachibana M, Kim S, Watarai M - J. Biomed. Sci. (2009)

Bottom Line: The recombinant TPR proteins were used for investigation of the effect of TPR proteins on bacterial uptake by TG cells and on pregnancy in mice.Bacterial TPR proteins bound to the C-terminal of Hsc70 through its EEVD motif and this binding inhibited bacterial uptake by TG cells.Our results demonstrate that surface located Hsc70 on TG cells mediates the uptake of pathogenic bacteria and proteins containing the TPR domain inhibit the function of Hsc70 by binding to its EEVD motif.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Veterinary Public Health, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan. kentaw@yamaguchi-u.ac.jp

ABSTRACT

Background: The uptake of abortion-inducing pathogens by trophoblast giant (TG) cells is a key event in infectious abortion. However, little is known about phagocytic functions of TG cells against the pathogens. Here we show that heat shock cognate protein 70 (Hsc70) contributes to bacterial uptake by TG cells and the EEVD motif of Hsc70 plays an important role in this.

Methods: Brucella abortus and Listeria monocytogenes were used as the bacterial antigen in this study. Recombinant proteins containing tetratricopeptide repeat (TPR) domains were constructed and confirmation of the binding capacity to Hsc70 was assessed by ELISA. The recombinant TPR proteins were used for investigation of the effect of TPR proteins on bacterial uptake by TG cells and on pregnancy in mice.

Results: The monoclonal antibody that inhibits bacterial uptake by TG cells reacted with the EEVD motif of Hsc70. Bacterial TPR proteins bound to the C-terminal of Hsc70 through its EEVD motif and this binding inhibited bacterial uptake by TG cells. Infectious abortion was also prevented by blocking the EEVD motif of Hsc70.

Conclusions: Our results demonstrate that surface located Hsc70 on TG cells mediates the uptake of pathogenic bacteria and proteins containing the TPR domain inhibit the function of Hsc70 by binding to its EEVD motif. These molecules may be useful in the development of methods for preventing infectious abortion.

Show MeSH

Related in: MedlinePlus

The domain structures of TPR proteins. TPR-Ba, TPR-Lm1, or TPR-Lm2 has six, five, or four TPR domains. TPR domain is indicated with striped bar. Domain search were performed using the SMART service http://smart.embl-heidelberg.de/.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2800845&req=5

Figure 3: The domain structures of TPR proteins. TPR-Ba, TPR-Lm1, or TPR-Lm2 has six, five, or four TPR domains. TPR domain is indicated with striped bar. Domain search were performed using the SMART service http://smart.embl-heidelberg.de/.

Mentions: Hsc70 has been reported to bind to proteins containing tetratricopeptide repeat (TPR) domains through its EEVD motif [17-19]. Therefore we hypothesized that bacterial proteins containing TPR domains could interact with Hsc70, and searched for such proteins in the genome database of B. abortus (GenBank AE017223, AE017224) and L. monocytogenes (GenBank AL591824). We found many TPR domain proteins in B. abortus and L. monocytogenes. In these TPR domain proteins, we selected a protein from B. abortus (BruAb2-0269, accession; YP_223064) and L. monocytogenes (Lmo-1510 and Lmo-2479, accession; NP_465035 and NP_466002) and designated them as TPR-Ba, TPR-Lm1, and TPR-Lm2, respectively. TPR-Ba and TPR-Lms (TPR-Lm1 and TPR-Lm2) contained six, five or four TPR domains (Fig. 3).


EEVD motif of heat shock cognate protein 70 contributes to bacterial uptake by trophoblast giant cells.

Watanabe K, Tachibana M, Kim S, Watarai M - J. Biomed. Sci. (2009)

The domain structures of TPR proteins. TPR-Ba, TPR-Lm1, or TPR-Lm2 has six, five, or four TPR domains. TPR domain is indicated with striped bar. Domain search were performed using the SMART service http://smart.embl-heidelberg.de/.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2800845&req=5

Figure 3: The domain structures of TPR proteins. TPR-Ba, TPR-Lm1, or TPR-Lm2 has six, five, or four TPR domains. TPR domain is indicated with striped bar. Domain search were performed using the SMART service http://smart.embl-heidelberg.de/.
Mentions: Hsc70 has been reported to bind to proteins containing tetratricopeptide repeat (TPR) domains through its EEVD motif [17-19]. Therefore we hypothesized that bacterial proteins containing TPR domains could interact with Hsc70, and searched for such proteins in the genome database of B. abortus (GenBank AE017223, AE017224) and L. monocytogenes (GenBank AL591824). We found many TPR domain proteins in B. abortus and L. monocytogenes. In these TPR domain proteins, we selected a protein from B. abortus (BruAb2-0269, accession; YP_223064) and L. monocytogenes (Lmo-1510 and Lmo-2479, accession; NP_465035 and NP_466002) and designated them as TPR-Ba, TPR-Lm1, and TPR-Lm2, respectively. TPR-Ba and TPR-Lms (TPR-Lm1 and TPR-Lm2) contained six, five or four TPR domains (Fig. 3).

Bottom Line: The recombinant TPR proteins were used for investigation of the effect of TPR proteins on bacterial uptake by TG cells and on pregnancy in mice.Bacterial TPR proteins bound to the C-terminal of Hsc70 through its EEVD motif and this binding inhibited bacterial uptake by TG cells.Our results demonstrate that surface located Hsc70 on TG cells mediates the uptake of pathogenic bacteria and proteins containing the TPR domain inhibit the function of Hsc70 by binding to its EEVD motif.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Veterinary Public Health, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan. kentaw@yamaguchi-u.ac.jp

ABSTRACT

Background: The uptake of abortion-inducing pathogens by trophoblast giant (TG) cells is a key event in infectious abortion. However, little is known about phagocytic functions of TG cells against the pathogens. Here we show that heat shock cognate protein 70 (Hsc70) contributes to bacterial uptake by TG cells and the EEVD motif of Hsc70 plays an important role in this.

Methods: Brucella abortus and Listeria monocytogenes were used as the bacterial antigen in this study. Recombinant proteins containing tetratricopeptide repeat (TPR) domains were constructed and confirmation of the binding capacity to Hsc70 was assessed by ELISA. The recombinant TPR proteins were used for investigation of the effect of TPR proteins on bacterial uptake by TG cells and on pregnancy in mice.

Results: The monoclonal antibody that inhibits bacterial uptake by TG cells reacted with the EEVD motif of Hsc70. Bacterial TPR proteins bound to the C-terminal of Hsc70 through its EEVD motif and this binding inhibited bacterial uptake by TG cells. Infectious abortion was also prevented by blocking the EEVD motif of Hsc70.

Conclusions: Our results demonstrate that surface located Hsc70 on TG cells mediates the uptake of pathogenic bacteria and proteins containing the TPR domain inhibit the function of Hsc70 by binding to its EEVD motif. These molecules may be useful in the development of methods for preventing infectious abortion.

Show MeSH
Related in: MedlinePlus