Adhesion characteristics of Listeria adhesion protein (LAP)-expressing Escherichia coli to Caco-2 cells and of recombinant LAP to eukaryotic receptor Hsp60 as examined in a surface plasmon resonance sensor.
Listeria adhesion protein (LAP) is an important adhesion factor in Listeria monocytogenes and interacts with its cognate receptor, mammalian heat shock protein 60 (Hsp60). The genetic identity of LAP was determined to be alcohol acetaldehyde dehydrogenase (Aad). A recombinant Escherichia coli strain expressing aad confirmed the involvement of Aad in adhesion to Caco-2 cells. Binding kinetics (ka) of recombinant LAP (rLAP) to Hsp60 was examined in a surface plasmon resonance sensor and was determined to be 5.35 x 10(8) M(-1) s(-1) and it was equivalent to the binding of anti-Hsp60 antibody (ka = 2.15 x 10(9) M(-1) s(-1)) to Hsp60. In contrast, Internalin B, an adhesion/invasion protein from L. monocytogenes, used as a control, had binding kinetics (ka) of only 2.9 x 10(6) M(-1) s(-1). The KD value of rLAP was 1.68 x 10(-8) M, which was significantly lower than Internalin B (KD = 6.5 x 10(-4) M). These results suggest that Hsp60 has significantly higher avidity for anti-Hsp60 antibody and LAP than Internalin B. In summary, LAP is identified as an alcohol acetaldehyde dehydrogenase and binding of recombinant E. coli to Caco-2 cells or rLAP to Hsp60 protein was found to be highly specific.
Kim
Kwang-Pyo
KP
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Jagadeesan
Balamurugan
B
Burkholder
Kristin M
KM
Jaradat
Ziad W
ZW
Wampler
Jennifer L
JL
Lathrop
Amanda A
AA
Morgan
Mark T
MT
Bhunia
Arun K
AK
D018829:Adhesins, Bacterial / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D018834:Chaperonin 60 / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000235:genetics; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance
D018829:Adhesins, Bacterial; D018834:Chaperonin 60; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase