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De novo pathway is an active metabolic pathway of cysteine synthesis in Haemonchus contortus

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Title De novo pathway is an active metabolic pathway of cysteine synthesis in Haemonchus contortus
 
Creator Vinod Kadam
Vijay Kumar Saxena
G.V. Vedamurthy
C.P. Swarnkar
Suneel Kumar Onteru
Haseen Ahmed
Raghvendar Singh
 
Subject Cysteine; Cysteine synthase; Cysteine proteases; de novo pathway; Haemonchus contortus
 
Description Not Available
Haemonchus contortus, commonly known as Barber's pole worm, is an economically important gastrointestinal nematode of sheep and goats especially in tropical and sub-tropical regions of the world. Cysteine synthesis is a very important metabolic pathway for the parasite, however the functional aspects of cysteine synthesis in parasite are largely unknown. The key question which we have investigated in the study is; whether the parasite uses a de novo pathway of cysteine synthesis, which is unknown in multicellular organisms of the animal kingdom and known to be absent in mammals. Directional cloning of the cysteine synthase (CS) gene was done in pET303 champion vector using restriction sites XbaI and XhoI. The CS gene of the H.contortus was closely related to CS-A protein of Oesophagostomum dentatum and a hypothetical protein of Ancylostoma ceylanicum. Recombinant protein of the H contortus CS (rHC-CS) gene was expressed using pET303 vector in pLysS BL21 strain of E.coli and subsequently purified by Ni-NTA affinity chromatography. Western blot using anti-His tag antibody confirmed the presence of rHC-CS. Biochemical assay, FTIR and enzyme kinetics studies revealed that rHC-CS used O-acetyl serine as substrate to produce cysteine using de novo pathway and CS activity was also confirmed with the homogenate of H.contortus. Upregulation of CS transcripts in the adult and its downregulation in the L3 larval stage suggests that de novo pathway contributes to the cysteine requirement of mature H.contortus. It is concluded that de novo pathway is an active metabolic pathway in H.contortus.
Not Available
 
Date 2021-07-30T05:58:27Z
2021-07-30T05:58:27Z
2021-05-31
 
Type Research Paper
 
Identifier Saxena, V. K., Vedamurthy, G. V., Swarnkar, C. P., Kadam, V., Onteru, S. K., Ahmad, H., & Singh, R. (2021). De novo pathway is an active metabolic pathway of cysteine synthesis in Haemonchus contortus. Biochimie.
0300-9084
http://krishi.icar.gov.in/jspui/handle/123456789/51432
 
Language English
 
Relation Not Available;
 
Publisher Elsevier