Molecular Mechanism for the Thermo-Sensitive Phenotype of CHO-MT58 Cell Line Harbouring a Mutant CTP : Phosphocholine Cytidylyltransferase
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Tartalom: http://real.mtak.hu/24974/
Archívum: MTA Könyvtár
Gyűjtemény: Status = Published


Type = Article
Cím:
Molecular Mechanism for the Thermo-Sensitive Phenotype of CHO-MT58 Cell Line Harbouring a Mutant CTP : Phosphocholine Cytidylyltransferase
Létrehozó:
Marton, Lívia
Nagy, Gergely N.
Ozohanics, Olivér
Lábas, Anikó
Krámos, Balázs
Oláh, Julianna
Vékey, Károly
Vértessy, Beáta G.
Dátum:
2015
Téma:
Q1 Science (General) / természettudomány általában
T2 Technology (General) / műszaki tudományok általában
Tartalmi leírás:
Control and elimination of malaria still represents a major public health challenge. Emerging parasite resistance to current therapies urges development of antimalarials with novel mechanism of action. Phospholipid biosynthesis of the Plasmodium parasite has been validated as promising candidate antimalarial target. The most prevalent de novo pathway for synthesis of phosphatidylcholine is the Kennedy pathway. Its regulatory and often also rate limiting step is catalyzed by CTP:phosphocholine cytidylyltransferase (CCT). The CHO-MT58 cell line expresses a mutant variant of CCT, and displays a thermo-sensitive phenotype. At non-permissive temperature (40 degrees C), the endogenous CCT activity decreases dramatically, blocking membrane synthesis and ultimately leading to apoptosis. In the present study we investigated the impact of the analogous mutation in a catalytic domain construct of Plasmodium falciparum CCT in order to explore the underlying molecular mechanism that explains this phenotype. We used temperature dependent enzyme activity measurements and modeling to investigate the functionality of the mutant enzyme. Furthermore, MS measurements were performed to determine the oligomerization state of the protein, and MD simulations to assess the inter-subunit interactions in the dimer. Our results demonstrate that the R681H mutation does not directly influence enzyme catalytic activity. Instead, it provokes increased heat-sensitivity by destabilizing the CCT dimer. This can possibly explain the significance of the PfCCT pseudoheterodimer organization in ensuring proper enzymatic function. This also provide an explanation for the observed thermo-sensitive phenotype of CHO-MT58 cell line.
Típus:
Article
PeerReviewed
info:eu-repo/semantics/article
Formátum:
text
Azonosító:
Marton, Lívia and Nagy, Gergely N. and Ozohanics, Olivér and Lábas, Anikó and Krámos, Balázs and Oláh, Julianna and Vékey, Károly and Vértessy, Beáta G. (2015) Molecular Mechanism for the Thermo-Sensitive Phenotype of CHO-MT58 Cell Line Harbouring a Mutant CTP : Phosphocholine Cytidylyltransferase. PLOS ONE, 10 (6). e0129632. ISSN 1932-6203
Kapcsolat:
Contract No 283570