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Chemotactic Activity of Cyclophilin A in the Skin Mucus of Yellow Catfish ( Pelteobagrus fulvidraco ) and Its Active Site for Chemotaxis

View Article: PubMed Central - PubMed

ABSTRACT

Fish skin mucus is a dynamic barrier for invading pathogens with a variety of anti-microbial enzymes, including cyclophilin A (CypA), a multi-functional protein with peptidyl-prolyl cis/trans isomerase (PPIase) activity. Beside various other immunological functions, CypA induces leucocytes migration in vitro in teleost. In the current study, we have discovered several novel immune-relevant proteins in yellow catfish skin mucus by mass spectrometry (MS). The CypA present among them was further detected by Western blot. Moreover, the CypA present in the skin mucus displayed strong chemotactic activity for yellow catfish leucocytes. Interestingly, asparagine (like arginine in mammals) at position 69 was the critical site in yellow catfish CypA involved in leucocyte attraction. These novel efforts do not only highlight the enzymatic texture of skin mucus, but signify CypA to be targeted for anti-inflammatory therapeutics.

No MeSH data available.


The expression and purification of GST-CypA. The CypA protein of yellow catfish was GST-fused expressed, purified and confirmed through SDS-PAGE, where the M represents the protein marker, Lane 1 represents CypAwt and Lane 2 represents CypAmt in this figure.
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ijms-17-01422-f003: The expression and purification of GST-CypA. The CypA protein of yellow catfish was GST-fused expressed, purified and confirmed through SDS-PAGE, where the M represents the protein marker, Lane 1 represents CypAwt and Lane 2 represents CypAmt in this figure.

Mentions: The wild-type and mutant-type CypA proteins (termed as CypAwt and CypAmt) were purified after expression and are shown in Figure 3. Both of the GST-fused proteins have an equal size of 43 kDa, and their chemotactic activity is depicted in Figure 4. The chemotactic index was about 3.08 in wild-type, while it was one in CypAmt, which is significantly lower than wild-type. This significant lower chemotactic index in the CypAmt confirms that Asn69 is responsible for chemotaxis.


Chemotactic Activity of Cyclophilin A in the Skin Mucus of Yellow Catfish ( Pelteobagrus fulvidraco ) and Its Active Site for Chemotaxis
The expression and purification of GST-CypA. The CypA protein of yellow catfish was GST-fused expressed, purified and confirmed through SDS-PAGE, where the M represents the protein marker, Lane 1 represents CypAwt and Lane 2 represents CypAmt in this figure.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037701&req=5

ijms-17-01422-f003: The expression and purification of GST-CypA. The CypA protein of yellow catfish was GST-fused expressed, purified and confirmed through SDS-PAGE, where the M represents the protein marker, Lane 1 represents CypAwt and Lane 2 represents CypAmt in this figure.
Mentions: The wild-type and mutant-type CypA proteins (termed as CypAwt and CypAmt) were purified after expression and are shown in Figure 3. Both of the GST-fused proteins have an equal size of 43 kDa, and their chemotactic activity is depicted in Figure 4. The chemotactic index was about 3.08 in wild-type, while it was one in CypAmt, which is significantly lower than wild-type. This significant lower chemotactic index in the CypAmt confirms that Asn69 is responsible for chemotaxis.

View Article: PubMed Central - PubMed

ABSTRACT

Fish skin mucus is a dynamic barrier for invading pathogens with a variety of anti-microbial enzymes, including cyclophilin A (CypA), a multi-functional protein with peptidyl-prolyl cis/trans isomerase (PPIase) activity. Beside various other immunological functions, CypA induces leucocytes migration in vitro in teleost. In the current study, we have discovered several novel immune-relevant proteins in yellow catfish skin mucus by mass spectrometry (MS). The CypA present among them was further detected by Western blot. Moreover, the CypA present in the skin mucus displayed strong chemotactic activity for yellow catfish leucocytes. Interestingly, asparagine (like arginine in mammals) at position 69 was the critical site in yellow catfish CypA involved in leucocyte attraction. These novel efforts do not only highlight the enzymatic texture of skin mucus, but signify CypA to be targeted for anti-inflammatory therapeutics.

No MeSH data available.