Essay on Lectin Agglutination

1477 Words6 Pages
Dwayne Hill.
BC34M
November 30, 2007
Lab Partner: Sasha-Kaye Graham
Experiment 6 : Lectin Agglutination
Aim:
To study the agglutination pattern of different strains of Psuedomonas aeruginosa to different sialic acid-binding lectins.
Absract:
The cell membrane of two strains of Psuedomonas aeruginosa, P22 and P201 were treated the sialic acid lectins SNA I, WGA, MAL and HAA to confirm the presence of sialic acid carbohydrates in the cell surface membranes of these strains. This was done by using two sets of samples-one treated with trypsin and the other without. The sample treated with trypsin showed agglutination for all lectins while the untreated sample showed little to no agglutination. A control was also used with the cells
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aeruginosa.
P.aeruginosa
Control
SNA
WGA
MAL
HAA
P22 (treated)
0
+++
+++
++
+++
P201 (treated)
0
+++
++
+
+++
P22 (untreated)
0
0
0
0
++
P201 (untreated)
0
0
0
0
0

0, no agglutination
+, weak agglutination
++, moderate agglutination
+++, strong agglutination
Questions:
1.It is important to study sialic acid binding lectins because their glyco-conjugates on the cell membrane of microorganisms are usually associated with virulence factors that cause disease eg. Streptococcus suis which has a high sialic acid content on its cell membrane causes infections in pigs and humans resulting in cases of meningitis and endocartis. Sialic acid is also associated with P. aeruginosa and the virulence it normally induces. Other lectins that are also of importance in microbiology and/or cell biology are mannose binding lectins which are also associated with virulence.
2.The agglutination is based on binding of the sialic acid to the cell membrane. To confirm that sialic acid carbohyrates are responsible for the binding the cells can be stripped of sialic acid and the experiment repeated.
The cells are treated with sialidase from C. Perfringens to remove sialic acid carbohydrates from the cell surface membrane.
Cell are given a 6 hour growth period
Cells are then centrifuged and re-suspended in sialidase and buffer and incubated to remove any sialic acid carbohydrates.
Cells are then washed in PBS

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