TY - CHAP
T1 - Detection of SUMOylation in Plasmodium falciparum
AU - Reiter, Katherine H.
AU - Matunis, Michael J.
N1 - Funding Information:
This work was supported by the National Institutes of Health (GM060980 to MJM) and the Sommer Scholar’s Program (KHR).
Publisher Copyright:
© Springer Science+Business Media New York 2016.
PY - 2016/9/1
Y1 - 2016/9/1
N2 - Reversible protein modification by small ubiquitin-related modifiers (SUMOs) regulates many cellular processes, including transcription, protein quality control, cell division, and oxidative stress. SUMOylation is therefore essential for normal cell function and represents a potentially valuable target for the development of inhibitors of pathogenic eukaryotic organisms, including the malaria parasite, Plasmodium falciparum (Pf). The specific and essential functions of SUMOylation in Pf, however, remain largely uncharacterized. The further development of antimalarial drugs targeting SUMOylation would benefit significantly from a more detailed understanding of its functions and regulation during the parasite life cycle. The recent development of antibodies specific for Pf SUMO provides a valuable tool to study the functions and regulation of SUMOylation. In preliminary studies, we have used immunoblot analysis to demonstrate that SUMOylation levels vary significantly in parasites during different stages of the red blood cell cycle and also in response to oxidative stress. Owing to the dynamic nature of SUMOylation and to the robust activity of SUMO isopeptidases, analysis of SUMOylation in cultured Pf parasites requires a number of precautions during parasite purification and lysis. Here, we outline methods for preserving SUMO conjugates during isolation of Pf parasites from human red blood cell cultures, and for their detection by immunoblot analysis using Pf SUMO-specific antibodies.
AB - Reversible protein modification by small ubiquitin-related modifiers (SUMOs) regulates many cellular processes, including transcription, protein quality control, cell division, and oxidative stress. SUMOylation is therefore essential for normal cell function and represents a potentially valuable target for the development of inhibitors of pathogenic eukaryotic organisms, including the malaria parasite, Plasmodium falciparum (Pf). The specific and essential functions of SUMOylation in Pf, however, remain largely uncharacterized. The further development of antimalarial drugs targeting SUMOylation would benefit significantly from a more detailed understanding of its functions and regulation during the parasite life cycle. The recent development of antibodies specific for Pf SUMO provides a valuable tool to study the functions and regulation of SUMOylation. In preliminary studies, we have used immunoblot analysis to demonstrate that SUMOylation levels vary significantly in parasites during different stages of the red blood cell cycle and also in response to oxidative stress. Owing to the dynamic nature of SUMOylation and to the robust activity of SUMO isopeptidases, analysis of SUMOylation in cultured Pf parasites requires a number of precautions during parasite purification and lysis. Here, we outline methods for preserving SUMO conjugates during isolation of Pf parasites from human red blood cell cultures, and for their detection by immunoblot analysis using Pf SUMO-specific antibodies.
KW - Immunoblotting
KW - Malaria
KW - Plasmodium falciparum
KW - Posttranslational modification
KW - SUMO
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U2 - 10.1007/978-1-4939-6358-4_19
DO - 10.1007/978-1-4939-6358-4_19
M3 - Chapter
C2 - 27631812
AN - SCOPUS:84987723145
T3 - Methods in Molecular Biology
SP - 283
EP - 290
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -