NDLI: Partial characterization of Acanthamoeba castellanii (T4 genotype) DNase activity

Content Provider	Springer Nature Link
Author	Iqbal, Junaid Panjwani, Shamvil Siddiqui, Ruqaiyyah Khan, Naveed Ahmed
Copyright Year	2014
Abstract	The deoxyribonuclease (DNase) activities of Acanthamoeba castellanii belonging to the T4 genotype were investigated. Using zymographic assays, the DNase activities had approximate molecular masses of 25 and 35 kDa. A. castellanii DNases exhibited activity at wide-ranging temperature of up to 60 °C and at pH ranging from 4 to 9. The DNases activities were unaffected by proteinase-K treatment, divalent cations such as Ca++, Cu++, Mg++, and Zn++, or divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) or sodium dodecyl sulfate (SDS). The non-reliance on divalent cations and homology data suggests that A. castellanii DNases belong to the class of eukaryotic lysosomal DNase II but exhibit robust properties. The DNases activity in A. castellanii interfered with the genomic DNA extraction. Extraction methods involving EDTA, SDS, and proteinase-K resulted in low yield of genomic DNA. On the other hand, these methods resulted in high yield of genomic DNA from human cells suggesting the robust nature of A. castellanii DNases that are unaffected by reagents normally used in blocking eukaryotic DNases. In contrast, the use of chaotropic agent such as guanidine thiocyanate improved the yield of genomic DNA from A. castellanii cells significantly. Further purification and characterization of Acanthamoeba DNases is needed to study their non-classic distinct properties and to determine their role in the biology, cellular differentiation, cell cycle progression, and arrest of Acanthamoeba.
Starting Page	457
Ending Page	463
Page Count	7
File Format	PDF
ISSN	09320113
Journal	Parasitology Research
Volume Number	114
Issue Number	2
e-ISSN	14321955
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-10-31
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	DNases Acanthamoeba castellanii Zymography Life cycle Medical Microbiology Microbiology Immunology
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Veterinary Parasitology Insect Science

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