TY - JOUR
T1 - Direct evidence for the atovaquone action on the Plasmodium cytochrome bc1 complex
AU - Siregar, Josephine E.
AU - Kurisu, Genji
AU - Kobayashi, Tamaki
AU - Matsuzaki, Motomichi
AU - Sakamoto, Kimitoshi
AU - Mi-ichi, Fumika
AU - Watanabe, Yoh ichi
AU - Hirai, Makoto
AU - Matsuoka, Hiroyuki
AU - Syafruddin, Din
AU - Marzuki, Sangkot
AU - Kita, Kiyoshi
N1 - Funding Information:
This paper is dedicated to Professor Kazuyuki Tanabe who has passed away on August 12, 2013. He has been a leading scientist in the field of malaria research and always encouraging us. This work was supported initially by a grant from the Indonesian government through the Ministry of Research and Technology and subsequently from the Japan Society for the Promotion of Sciences (JSPS) to JES, through JSPS fellowship for Ph.D. RONPAKU program (to JES), and was supported by Grant-in-aid for Scientific Research ( 26253025 ) to KK from the Japanese Society for the Promotion of Science . We also acknowledge the support from the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries and Food Industry and JST / JICA , SATREPS (Science and Technology Research Partnership for Sustainable Development) ( 10000284 ) to KK.
Publisher Copyright:
© 2014.
PY - 2015/6/1
Y1 - 2015/6/1
N2 - Atovaquone, a coenzyme Q analogue has been indicated to specifically target the cytochrome bc1 complex of the mitochondrial respiratory chain in the malarial parasite and other protozoan. Various mutations in the quinone binding site of the cytochrome b gene of Plasmodium spp. such as M133I, L144S, L271V, K272R, Y268C, Y268S, Y268N, and V284F are suggesting to associate with resistance to atovaquone. There is no direct evidence of relation between the mutations and resistance to atovaquone in Plasmodium parasite that has been available. Technical difficulties in isolating active assayable mitochondria in the malarial parasite hinder us to obtain direct biochemical evidence to support the relation between the mutations and drug resistance.The establishment of a mitochondrial isolation method for the malaria parasite has allowed us to test the degree of resistance of Plasmodium berghei isolates to atovaquone directly. We have tested the activity of dihydroorotate (DHO)-cytochrome c reductase in various P. berghei atovaquone resistant clones in the presence of a wide concentration range of atovaquone. Our results show the IC50 of P. berghei atovaquone resistant clones is much higher (1.5 up to 40nM) in comparison to the atovaquone sensitive clones (0.132-0.465nM). The highest IC50 was revealed in clones carrying Y268C and Y268N mutations (which play an important role in atovaquone resistance in Plasmodium falciparum), with an approximately 100-fold increase. The findings indicate the importance of the mutation in the quinone binding site of the cytochrome b gene and that provide a direct evidence for the atovaquone inhibitory mechanism in the cytochrome bc1 complex of the parasite.
AB - Atovaquone, a coenzyme Q analogue has been indicated to specifically target the cytochrome bc1 complex of the mitochondrial respiratory chain in the malarial parasite and other protozoan. Various mutations in the quinone binding site of the cytochrome b gene of Plasmodium spp. such as M133I, L144S, L271V, K272R, Y268C, Y268S, Y268N, and V284F are suggesting to associate with resistance to atovaquone. There is no direct evidence of relation between the mutations and resistance to atovaquone in Plasmodium parasite that has been available. Technical difficulties in isolating active assayable mitochondria in the malarial parasite hinder us to obtain direct biochemical evidence to support the relation between the mutations and drug resistance.The establishment of a mitochondrial isolation method for the malaria parasite has allowed us to test the degree of resistance of Plasmodium berghei isolates to atovaquone directly. We have tested the activity of dihydroorotate (DHO)-cytochrome c reductase in various P. berghei atovaquone resistant clones in the presence of a wide concentration range of atovaquone. Our results show the IC50 of P. berghei atovaquone resistant clones is much higher (1.5 up to 40nM) in comparison to the atovaquone sensitive clones (0.132-0.465nM). The highest IC50 was revealed in clones carrying Y268C and Y268N mutations (which play an important role in atovaquone resistance in Plasmodium falciparum), with an approximately 100-fold increase. The findings indicate the importance of the mutation in the quinone binding site of the cytochrome b gene and that provide a direct evidence for the atovaquone inhibitory mechanism in the cytochrome bc1 complex of the parasite.
KW - Atovaquone-resistance
KW - Cytochrome bc complex
KW - Malaria
KW - Plasmodium
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U2 - 10.1016/j.parint.2014.09.011
DO - 10.1016/j.parint.2014.09.011
M3 - Article
C2 - 25264100
AN - SCOPUS:84924404654
SN - 1383-5769
VL - 64
SP - 295
EP - 300
JO - Parasitology International
JF - Parasitology International
IS - 3
ER -