International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-

8,2beta-0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence
Protein of Uropathogenic Escherichia Coli

Abhilash M.

Department of Biotechnology, The Oxford college of Engineering, Bangalore, India

Corresponding author email [email protected]

ABSTRACT

Fimbrial adhesion is a Virulence Determinant which is classified under Adhesins category of virulence
factor of uropathogenic Escherichia coli. Afimbrial adhesin Protein with swissprot accession number P12730,
of length 181 amino was selected for modeling using Bioinformatics tools. Modelled protein has been
submitted to protein model database and has been assigned an accession number of PM0075877. Docking
analysis of Epicatechin(4beta-8,2beta-0-7), a Proanthocyanin from cranberry against modelled fimbrial
adhesion was carried out using hex docking tool. Some of the commonly used antibiotics to treat urinary tract
infections caused by Uropathogenic E.coli which includes Oflaxacin, sulfamethoxazole, Trimethoprim were
subjected to docking analysis for comparative studies.

KEY WORDS: Uropathogenic E-coli, Fimbrial adhesin, Epicatechin, Docking analysis.

INTRODUCTION of bacteria in the urinary tract1. This disease is

associated with many virulence factors present in
UPEC 2. Uropathogenic Escherichia coli (UPEC) are
Urinary tract infection is one of the most important
the most important group of microorganisms
causes of morbidity and mortality. E.coli is the most
responsible for urinary tract infection. UPEC differ
frequent urinary pathogen isolated from 50% - 90% of
from non-pathogenic E. coli and from other E. coli
all uncomplicated urinary tract infections . The
pathotypes by the production of specific virulence
urinary tract infection (UTI) is defined by the presence
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 International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-8,2beta-

0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence Protein of
Uropathogenic Escherichia Coli
factors, which enable the bacteria to adhere to adherence of E.coli strains is specifically inhibited
uroepithelial cells and to establish urinary tract by D-mannose (Man) and methyl-a- D-mannoside,
infections 3. UPEC can evade the body's innate and is thus called mannose specific or mannose
immune defenses by invading superficial umbrella sensitive11 . It has been shown that type 1 pili bind to
cells to form intracellular bacterial communities guinea pig erythrocytes , as well as to human buccal
(IBCs) 4 . The adherence of Escherichia coli to cells and yeast cells12.
uroepithelial cells is an important event in the
pathogenesis of urinary tract infections5. Although Use of Cranberries- Vaccinium macrocarpon to treat
afimbrial adhesins have been identified6, bacterium- UTI is well documented13. Cranberry juice acts by
to-host cell attachment is usually mediated by preventing adhesion which presumably helps urinary
fimbriae (pili). flushing of the causative bacteria, preventing their
Fimbriae are used by bacteria to adhere to one colonization of the urinary tract. This effect is
another and to adhere to animal cells, and some achieved by inhibiting the infecting bacteria,
inanimate objects7 . Their presence greatly enhances Escherichia coli, from adhering to uroepithelial
the bacteria's ability to attach to the host and cause cells14.
disease8 . For Escherichia coli strains of various
origins, only some of the epithelial receptors are now Cranberries contain a unique polymeric compound
known at the molecular level, and the concerned known as proanthocyanidin15. Proanthocyanidin
adhesins can be placed in three distinct groups with shows a very strong inhibitory activity against
regard to their carbohydrate specificities9 . mannose-resistant adhesins produced by urinary
isolates of E. coli16. Cranberries inhibit bacterial
I. The first group includes adhesins that recognize adhesion to the uroepithelium, preventing bacterial
digalactoside moieties like the P fimbriae of human multiplication and the ensuing bladder infection17.
pyelonephritic E.coli strains specific for the P blood And hence these compounds can be used to treat
group antigens. Urinary tract infections.
II. The second group comprises adhesins recognizing
sialylated glycoconjugates and includes the M MATERIALS AND METHODS METHODS
fimbriae of human pyelonephritic E.coli strains
specific for the blood group M determinant of
Protein with Swiss-Prot primary accession number
glycophorin A10.
P12730, of length 181 amino
III. The third group includes type 1 fimbriae that are
acids was selected for modelling of Afimbrial adhesin
expressed by many strains of nonpathogenic and
AFA-I.
pathogenic E.coli and which are isolated, for
instance, from patients with diarrheal diseases or
Modelling
urinary tract infections or from animals with
diarrhea. When mediated by type 1 pili, the bacterial
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 International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-8,2beta-

0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence Protein of
Uropathogenic Escherichia Coli
Modelling of target protein was carried out using and the template sequence is provided.The sequence
Swiss-PdbViewer (or SPDBV). Modelling of protein alignment and template structure are then used to
refers to constructing an atomic-resolution model of produce a structural model of the target.
the target protein from its amino acid sequence and an Inhibitor Molecule
experimental three-dimensional structure of a related Epicatechin(4beta-8,2beta-0-7)epicatechin a
homologous protein template. The template with PDB Proanthocyanidin from cranberry was selected as
accession number 2JTYA was selected for modelling. lead molecule for Insilco analysis of its inhibition
The amino acid sequence of the target to be modeled activity against modelled fimbrial adhesion.

Figure 1: Epicatechin(4beta-8,2beta-0-7)epicatechin a Proanthocyanidin from cranberry

Lead Validation was also used to visualize Interaction of

Epicatechin-(4beta 2)-phloroglucinol with
OSIRIS Property Xplorer was used to validate modelled fimbrial adhesion protein. Some of the
the lead molecule which has inhibitory effect on commonly used antibiotics to treat urinary tract
the modeled target protein. infections caused by Uropathogenic E.coli which
includes Oflaxacin, sulfamethoxazole,
Docking Studies Trimethoprim were subjected to docking analysis
Hex 5.1 tool was used for Docking studies. for comparative studies using Hex 5.1.
Analysis of Epicatechin(4beta-8,2beta-0-
7)epicatechin a Proanthocyanidin from cranberry RESULTS AND ISCUSSION
ISCUSSION
as an inhibitor against modelled fimbrial adhesion
Modelling
was carried out using Hex 5.1. Further Hex 5.1

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 International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-

8,2beta-0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence
Protein of Uropathogenic Escherichia Coli
Modelling of fimbrial adhesin protein was done an accession number PM0075877.This accession
using SPDBV tool. Modelled protein structure number can be used to retreive the submitted
was submitted to protein model database. proteinstructure.
Submitted modelled structure has been assigned

Figure 2: Structure of modelled fimbrial adhesin protein

Lead validation
Properties such as mutagenecity, irritant, compound against the modeled target protein was
tumorogenic ,drug likeliness, of the lead molecule carried out using OSIRIS Property Xplorer.
were studied and validation of this inhibitory

Figure 3: Result of validation assay

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 International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-8,2beta-0-

7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence Protein of
Uropathogenic Escherichia Coli

It is observed that Epicatechin(4beta-8,2beta-0-7)epicatechin clears mutagenecity, druglikeness , tumorogenic,

irritant assays. Hence, can be considered as potential inhibitor for further analysis.

Docking analysis
Docking analysis of Epicatechin-(4beta 2)-commonly used antibiotics such as Oflaxacin,
phloroglucinol a Proanthocyanidin from cranberry asSulfamethoxazole and Trimethoprim, to treat Urinary
an inhibitor against modelled fimbrial adhesion wasTract Infections were subjected to Docking Analysis to
carried out using Hex 5.1.Interaction of Epicatechin-compare them with the lead molecule. The Binding
(4beta 2)-phloroglucinol with the modelled target proteinEnergy values obtained for the lead molecule and these
and its E value is shown in figure 2. Some of the mostantibiotics are shown in table 1.

Figure 4: Interaction of Epicatechin-(4beta 2)-phloroglucinol a Proanthocyanidin from cranberry with

Modelled fimbrial adhesion virulence protein.

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 International Journal of Pharma and Bio Sciences V1(1),2010

Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-

8,2beta-0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence
Protein of Uropathogenic Escherichia Coli
E Value: -243.59 k cal/mol

Table 1: Comparison of E values between Existing Antibiotics to treat UTI and the Lead molecule
∆G
Sl. No. NAME OF THE COMPOUND
kcal/mol
Lead Molecule
-243.59
Epicatechin-(4beta 2)-phloroglucinol
Commonly used Antibiotics
1 Ofloxacin -190.88
2 Trimethoprim -183.28
3 Sulfamethoxazole -170.00

CONCLUSION Raju,Executive director of Children’s education trust,

Dr.T.Krishnan Principal, Dr. Kusum Paul of The
Since its reported that Uropathogenic E Coli have oxford college of Engineering, Bangalore for their
developed antibiotic resistance(Gupta et al 2002), support and encouragement.
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Insilico Analysis of Cranberry Proanthocyanidin Epicatechin(4beta-8,2beta-

0-7) As an Inhibitor for Modelled Afimbrial Adhesin Virulence Protein of
Uropathogenic Escherichia Coli
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