Determining the substrates for MAP kinases in Leishmania mexicana

Leishmaniasis is a protozoan infection caused by Leishmania species, members of the parasitic trypanosomtidae family. Leishmania mexicana is a 'New World' species causing cutaneous leishmaniasis. Resistance to the current drug treatments has been observed for Leishmaniasis and with this resistance emerging, research into new drug targets is essential. Mitogen-activated protein (MAP) kinases present a potential target; fifteen different MAP kinases have been found in L. mexicana, some of which have proven to be essential at various stages in the Leishmania life cycle. This project focuses on identifying substrates for three of these kinases: LmxMPK2, LmxMPK3 and LmxMPK13, through the use of short GST-fusion peptides representing phosphorylation sites found in particular proteins by mass spectrometry. Seventeen potential substrates were found from the fifty GST-fusion peptides tested. One of these peptides, OSM3-like kinesin, was used for further study, because there appeared to be a clear link between the kinases and substrate all being involved in flagellar processes. The OSM3-like kinesin peptide was found to be a potential substrate of LmxMPK3 and LmxMPK13, two kinases known to be involved in flagellum length regulation. Proteins related to OSM3-like kinesin have been found to be involved in intra-flagellar transport. Localisation of OSM3 using a GFP-fusion construct showed that the protein localised to the flagellum in L. mexicana, indicating an in vivo interaction of the kinases and OSM3-like kinesin.

Name dc:type.qualificationname

mres

Level dc:type.qualificationlevel

masters-pg

Grantor dc:publisher.institution

University of Strathclyde

Year dc:date.issued

2014

Author dc:creator

• Emmerson, Josiah Samuel

DOI

10.48730/aq12-3p90

Identifier

T13718

OAI identifier oai:identifier

oai:strathclyde:ww72bb51z