Effect of chemical modifications on the K99 and K88ab fibrillar adhesins of Escherichia coli

Jacobs, A. A. C.; Mechelen, J.R. van; Graaf, Frits K. de

doi:10.1016/0167-4838(85)90326-7

Cited by 16 publications

(7 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Chemical modification of different amino acid species in the K88 adhesin has shown that arginine residues also may play a role in the receptor-binding event (11). The significance of this observation and the location of the respective arginine residues in the K88 primary structure are presently under investigation.…”

Section: Resultsmentioning

confidence: 95%

See 1 more Smart Citation

Role of phenylalanine 150 in the receptor-binding domain of the K88 fibrillar subunit

et al. 1987

View full text Add to dashboard Cite

Recently, we reported the isolation of three peptides, Ile-83-Ala-Phe-85, Ser-148-Leu-Phe-150, and Ala-156-Ile-Phe-158, derived from the K88 fibrillar subunit and found to inhibit the binding of K88 fibrillae to cavia erythrocytes or pig intestinal epithelial cells (A. A. C. Jacobs, J. Venema, R. Leeven, H. van Pelt-Heerschap, and F. K. de Graaf, J. Bacteriol. 169:735-741, 1987). The gene encoding the K88 fibrillar adhesin was modified by oligonucleotide-directed site-specific mutagenesis such that each of the phenylalanine residues at positions 85, 150, and 158 were replaced by serine. Replacement of phenylalanine 85 or 158 had no apparent effect on the biosynthesis of the fibrillae or on their adhesive capacity. In contrast, substitution of phenylalanine 150 with serine resulted in a dramatic decrease in adhesive capacity of the K88 fibrillae. Apparently, phenylalanine 150 plays an essential role in the interaction of the adhesin with receptor molecules present on eucaryotic cells.

show abstract

Section: Resultsmentioning

confidence: 95%

“…The structure of the K88 receptor(s) is unknown, but there are several indications that galactosyl residues are involved in the specific binding (5). Chemical modification studies implicated a role for arginine residues in the receptor-binding domain of the K88 adhesin, but the modified residues have not been located in the primary structure (11).…”

mentioning

confidence: 99%

Role of phenylalanine 150 in the receptor-binding domain of the K88 fibrillar subunit

et al. 1987

View full text Add to dashboard Cite

show abstract

“…In addition, the relatively high concentrations of tripeptide required to inhibit the hemagglutinating activity of K88ad fibrillae suggests that differences in receptor molecules exist in particular between the receptors for K88ad and for K88ab or -ac. Recent biochemical studies showed that modification of two arginine residues on an average of one adhesin molecule results in the loss of hemagglutinating activity of the K88ab fibrillae (11). This indicates that besides hydrophobic amino acid side chains charged residues also contribute to receptor binding.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of adhesive activity of K88 fibrillae by peptides derived from the K88 adhesin

Jacobs¹,

Venema²,

Leeven³

et al. 1987

J Bacteriol

View full text Add to dashboard Cite

A cyanogen bromide fragment derived from the K88ab adhesin inhibited the hemagglutinating activity of K88 fibrillae. Smaller fragments which inhibited the adherence of K88 fibrillae to erythrocytes or to intestinal epithelial cells were obtained by digestion of K88ab fibrillae with oa-chymotrypsin. Active peptides were isolated from the digestion mixture and identified as Ser-Leu-Phe and Ala-Ile-Phe. Both tripeptides correspond to the peptide stretches Ser-148-Leu-Phe-150 and Ala-156-Ile-Phe-158, respectively, which are part of conserved regions in the primary structure of the K88 variants ab, ac, and ad. The isolated tripeptides inhibited the hemagglutinating activity of purified K88 fibrillae in the 1 to 5 ,uM range, while adherence of the fibrillae to intestinal epithelial cell brush borders was inhibited in the 10 to 50 ,uM range. Furthermore, the tripeptides were capable of eluting attached bacteria from agglutinated erythrocytes. The inhibitory activity of the isolated peptides was confirmed by testing various synthetic peptides for their ability to inhibit the interaction of the different K88 variants with various species of erythrocytes. The significance of these findings for the localization of the receptor-binding domain is discussed.K88 fibrillae are nonflagellar, filamentous adhesins found on many enterotoxigenic Escherichia coli strains that cause neonatal diarrhea in pigs (4). They enable the bacteria to colonize the small intestinal epithelium, which is considered to be a prerequisite for the establishment of diarrheal disease. The K88 fibrillae consist of multimers of the K88 adhesin subunit with a molecular weight of 27,500 (4). Three serological variants of K88 fibrillae have been described and designated K88ab, K88ac, and K88ad (7). The primary structure of all three proteins has been determined, showing both conserved and variable regions (5,6,12 Purification of fibrillae. K88ab, K88ac, K88ad, F41, and K99 fibrillae were isolated and purified as described previously (10).Isolation of brush borders. Brush borders were prepared from the pig intestine as described by Middeldorp and Witholt (15) and stored in 50% glycerol at -20°C.Hemagglutination inhibition test. Suspensions of washed erythrocytes (2%) in PBSM (50 mM sodium phosphate [pH 7.3] containing 0.9% NaCl and 0.5% mannose) were mixed 1:1 with various amounts of peptide fractions in the same buffer. Subsequently, 50-pl portions of these suspensions were added to serial twofold dilutions (50 ,ul) of purified fibrillae in PBSM, using polystyrene microtiter trays with V-shaped cups. After 2 h of incubation at 4°C, the trays were examined. The initial concentrations of K88ab, -ac, and -ad fibrillae used were 50, 500, and 25 [Lg/ml, respectively

show abstract

“…To this end, the bacteria have regulatory systems at their disposal that enable them to recognize the environmental circumstances and to adapt fimbriae biosynthesis according to these conditions. In the case of F4 fimbriae, expression is optimal at a temperature of 37°C, a pH ranging from 6.5 to 8.0 and at the end of the exponential growth [Jacobs et al, 1985;Mooi and de Graaf, 1985;van Verseveld et al, 1985;Blomberg et al, 1991;Payne et al, 1993]. This regulation of fimbrial expression is mediated by several factors like global regulators, local regulators, crosstalk between fimbriae and posttranscriptional mechanisms.…”

Section: Regulation Of F4 Expressionmentioning

confidence: 99%

“…The biosynthesis of fimbriae is influenced by growth rate [Jacobs et al, 1985;van der Woude et al, 1990a;Blomberg et al, 1991Blomberg et al, , 1993 and environmental conditions such as carbon source [Shipley et al, 1978;Blomberg et al, 1991;White-Ziegler et al, 2000], temperature [Mooi et al, 1979;Göransson, 1984Göransson, , 1990Klemm, 1985;Mooi and de Graaf, 1985;Schmoll et al, 1990;van der Woude et al, 1990b;White-Ziegler et al, 1990Gally et al, 1993], pH [Schwan et al, 2002;van Verseveld et al, 1985;White-Ziegler et al, 2000], osmolarity [Spears et al, 1986;Göransson et al, 1990;White-Ziegler et al, 2000;Schwan et al, 2002], oxygen levels [White-Ziegler et al, 2000] and presence of alanine or leucine de Graaf and Mooi, 1986;Braaten et al, 1992;Gally et al, 1993]. Indeed, pathogenic bacteria only fully express virulence factors like fimbriae when the conditions are appropriate for adherence and subsequent colonization [Pourbakhsh et al, 1997;Zhao et al, 1997;Struve and Krogfelt, 1999].…”

Section: Regulation Of F4 Expressionmentioning

confidence: 99%

F4 Fimbriae Expressed by Porcine Enterotoxigenic <i>Escherichia coli</i>, an Example of an Eccentric Fimbrial System?

Verdonck

Cox²,

Goddeeris³

2004

Microb Physiol

View full text Add to dashboard Cite

An overwhelming number of infectious diseases in both humans and animals are initiated by bacterial adhesion to carbohydrate structures on a mucosal surface. Most bacterial pathogens mediate this adhesion by fimbriae or pili which contain an adhesive lectin subunit. The importance of fimbriae as virulence factors led to research elucidating the regulation of fimbrial expression and their molecular assembly process. This review provides an overview of the current knowledge of induction, expression and assembly of F4 (K88) fimbriae and discusses its unique as well as its identical characteristics compared to other intensively studied fimbriae or pili expressed by Escherichia coli.

show abstract

Effect of chemical modifications on the K99 and K88ab fibrillar adhesins of Escherichia coli

Cited by 16 publications

References 21 publications

Role of phenylalanine 150 in the receptor-binding domain of the K88 fibrillar subunit

Role of phenylalanine 150 in the receptor-binding domain of the K88 fibrillar subunit

Inhibition of adhesive activity of K88 fibrillae by peptides derived from the K88 adhesin

F4 Fimbriae Expressed by Porcine Enterotoxigenic <i>Escherichia coli</i>, an Example of an Eccentric Fimbrial System?

Contact Info

Product

Resources

About