Adhesion properties of Lactobacillus rhamnosus mucus-binding factor to mucin and extracellular matrix proteins

Abstract

We previously described potential probiotic Lactobacillus rhamnosus strains, isolated from fermented mare milk produced in Sumbawa Island, Indonesia, which showed high adhesion to porcine colonic mucin (PCM) and extracellular matrix (ECM) proteins. Recently, mucus-binding factor (MBF) was found in the GG strain of L. rhamnosus as a mucin-binding protein. In this study, we assessed the ability of recombinant MBF protein from the FSMM22 strain, one of the isolates of L. rhamnosus from fermented Sumbawa mare milk, to adhere to PCM and ECM proteins by overlay dot blot and Biacore assays. MBF bound to PCM, laminin, collagen IV, and fibronectin with submicromolar dissociation constants. Adhesion of the FSMM22 mbf mutant strain to PCM and ECM proteins was significantly less than that of the wild-type strain. Collectively, these results suggested that MBF contribute to L. rhamnosus host colonization via mucin and ECM protein binding.

Cell surface‐associated aggregation‐promoting factor from Lactobacillus gasseri SBT2055 facilitates host colonization and competitive exclusion of Campylobacter jejuni

“…Moreover, GST‐APF1 bound tightly to fibronectin and exhibited a K D value of 68.7 nM. Previous studies have shown high affinity to fibronectin of the S‐layer protein SlpA from L. brevis ATCC 8287 ( K D = 89.8 nM) (de Leeuw et al ., ), and of the mucus‐binding factor from L. rhamnosus FSMM22 ( K D = 190 nM) (Nishiyama et al ., ). Our findings suggest that APF1 may belong to a class of Lactobacillus adhesins that exhibit high affinity for fibronectin.…”

“…A similar interaction has been observed for the S‐layer protein CbsA from L. crispatus JCM 5810, which binds laminin, collagens and lipoteichoic acids (Antikainen et al ., ). Other studies have demonstrated the affinity of glutamine synthetase and glucose‐6‐phosphate isomerase from L. crispatus ST1 to laminin, collagen I and fibronectin (Kainulainen et al ., ), and that of mucus‐binding factor from L. rhamnosus strains GG and FSMM22 to laminin, collagen IV, fibronectin and mucin (Nishiyama et al ., ). Bacterial adhesion has been suggested to depend on cell surface hydrophobicity and/or electric charge (Schär‐Zammaretti et al ., ).…”

“…Primer pairs KN07/KN08 (for apf1 ) and KN09/KN10 (for apf2 ) (http://onlinelibrary.wiley.com/doi/10.1111/mmi.13153/suppinfo) were used to introduce blunt ends in the middle of the apf genes in pGEM‐T‐ apf1 and pGEM‐T‐ apf2 , respectively, by PCR. The pGEM‐T‐ cm r plasmid (Nishiyama et al ., ) was digested with Bam HI and Hind III to obtain a terminator‐less Cm‐resistance cassette, which was treated with the Klenow fragment (Takara Bio) and inserted into pGEM‐T‐ apf1 and pGEM‐T‐ apf2 . The resultant plasmids carrying the chloramphenicol resistance cassette in the middle of the apf genes in the same direction were selected and designated as pGEM‐T‐ apf1 :: cm r and pGEM‐T‐ apf2 :: cm r respectively.…”

Use of Atomic Force Microscopy to Study the Multi-Modular Interaction of Bacterial Adhesins to Mucins

“…Methodologies to screen for bacterial adhesion to mucins have previously employed thin layer chromatography overlay [26], enzyme-linked immunosorbent assay [15], micro-titre plate assays [8,25,27], surface plasmon resonance [16,28,29,30,31], fluorescence spectroscopy [20], mucin microarrays [32], flow cytometry [33], and cell-based assays [26,34,35,36]. However, due to the complexity and diversity of mucin glycosylation, these methods typically provide qualitative binding data indicating only presence or absence of interaction.…”

In vitroinvestigation of molecules involved inLactobacillus gasseriSBT2055 adhesion to host intestinal tract components

“…rhamnosus FSMM22 (Nishiyama et al . ). Two surface layer proteins are known Fn‐binding proteins: SlpA of Lactobacillus brevis ATCC 8287 (Hynönen et al .…”