Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
Melioidosis is a disease that infects humans and animals, and can be detrimental in humans. Mortality rate from melioidosis septic shock due to infection from Gram negative Burkholderia pseudomallei (B. pseudomallei) in endemic regions of Malaysia and Thailand remains high despite available antim...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English English |
| Published: |
Universiti Putra Malaysia
2019
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/72576/ http://irep.iium.edu.my/72576/ http://irep.iium.edu.my/72576/1/15%20JTAS-1245-2017.pdf http://irep.iium.edu.my/72576/7/72576_Partial%20Purification%20and%20Model%20Structure_scopus.pdf |
| Summary: | Melioidosis is a disease that infects humans and animals, and can be detrimental in
humans. Mortality rate from melioidosis septic shock due to infection from Gram
negative Burkholderia pseudomallei (B. pseudomallei) in endemic regions of Malaysia
and Thailand remains high despite available antimicrobial therapy. Multiple strategies are
employed to identify essential genes and drug targets in this bacterium to improve current
antimicrobial therapies. This is important as B. pseudomallei is intrinsically resistant
to many commonly used antibiotics. In this study, hypothetical genes predicted to be
essential for B. pseudomallei by transposon-directed insertion site sequencing (TraDIS)
technique were selected. One target gene, BPSL2774, has been successfully amplified and
cloned from genomic DNA of B. pseudomallei strain K96243. Glutathione S-transferase
(GST) affinity tag chromatography was performed for partial protein purification. The
target protein was successfully expressed in soluble form with satisfactory yield output.
Mass spectrometry analysis of 60 kDa
Coomassie-stained gel band confirmed the
presence of the soluble expressed taggedtarget protein, co-purified with Escherichia
coli chaperonin proteins, possibly due to
their interaction with the target protein.
BPSL2774 protein have considerable
homology to glycosyltransferase GTB
type superfamily and RfaB superfamily.
On the basis of this similarity, the three-dimensional structure of BPSL2774 has
been modelled and assessed by protein
model quality servers. Taking all the results
into account, the functional annotation of
BPSL2774 protein as a glycosyltransferase
is recommended, though future validation
from biochemical experiments will be
needed to support this. |
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