Characterization of the type 3 fimbriae with different MrkD adhesins: possible role of the MrkD containing an RGD motif |
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Authors: | Huang Ying-Jung Wu Chien-Chen Chen Mei-Chen Fung Chang-Phone Peng Hwei-Ling |
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Institution: | Department of Biological Science and Technology, National Chiao Tung University, Hsin Chu 30050, Taiwan, ROC. |
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Abstract: | Four novel mrkD alleles namely mrkD(V1), mrkD(V2), mrkD(V3), and mrkD(V4) were identified in seventeen Klebsiella pneumoniae meningitis strains using PCR-RFLP and sequence determination. Comparative analysis revealed a most variable region containing an RGD motif in the receptor domain of MrkD(V3). In order to determine if the sequence confers the K. pneumoniae mrkD(V3) the highest level of the fimbrial activity, a type 3 fimbriae display system was constructed in Escherichia coli. The E. coli JM109pmrkABCD(V3)F] displaying meshwork-like fimbriae also had the most fimbrial activity, supporting a possible role of the varied sequences. In a dose-dependent manner, the GRGDSP hexapeptide appeared to inhibit the adhesion of the E. coli JM109pmrkABCD(V3)F] to HCT-8, an ileocecal epithelial cell line. In addition, the adhesion activity was reduced by the addition of anti-alpha5beta1 integrin monoclonal antibody, indicating that the RGD containing region in MrkD(V3) is responsible for the binding of type 3 fimbriae to integrin. |
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Keywords: | Klebsiella pneumoniae Type 3 fimbriae mrkD PCR-RFLP RGD motif Integrin |
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