Cloning, expression, purification and crystallization of a novel GlcNAc metabolic protein, gig2 (duf1479) from pathogenic fungus Candida albicans
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Title |
Cloning, expression, purification and crystallization of a novel GlcNAc metabolic protein, gig2 (duf1479) from pathogenic fungus Candida albicans
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Creator |
Rani, Priya
Gautam, Gunjan Rao, Kongara Hanumantha Ghosh, Swagata Gourinath, Samudrala Dhar, Suman Kumar Datta, Asis |
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Subject |
N-acetylglucosamine (GlcNAc)
GlcNAc Inducible Gene 2 (GIG2) DUF (Domains of Unknown Function) family of proteins Crystallography |
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Description |
Accepted date: June 21, 2017
N-acetylglucosamine (GlcNAc), an alternative sugar, is emerging as an important molecule having a multifarious role in Candida albicans including a major role in signaling. GlcNAc Inducible Gene 2, GIG2 is one of the highly upregulated genes in GlcNAc grown cells in C. albicans. Our earlier studies show the involvement of Gig2 in the formation of N-acetylneuraminic (NANA) acid from GlcNAc-6-phosphate through an understudied route. The crystal structure of Gig2 would help us in determining the exact reaction that this enzyme catalyzes. Here the cloning, expression, purification and crystallization of this protein are reported along with preliminary X-ray crystallographic analysis at 2.4Å resolution. The crystal belonged to P2 1 space group, with unit cell parameters a=59.59, b= 54.43, c= 73.29Å; α = 90°, β = 102.7° and γ = 90°. The structure was solved using PDB ID 2CSG as a template which has only 27% identity. Molecular replacement yielded a solution with LLG score of 87. The structure is currently under further refinement. We thank Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Government of India for funding. We thank UGC-RNW, UGC-SAP, UGCUPOE-II, DST-FIST, and DST-PURSE for funding central instrumentation facility and for extending institutional funding. PR and GG thank UGC and CSIR for fellowship. We also thank Department of Biotechnology (DBT) and staffs of BM14, ESRF for helping us with the collection of high-resolution X-ray data. |
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Date |
2018-06-25T08:13:11Z
2018-06-25T08:13:11Z 2017 |
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Type |
Article
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Identifier |
Journal of Proteins and Proteomics 8(2): 127-132
0975-8151 http://223.31.159.10:8080/jspui/handle/123456789/865 http://jpp.org.in/index.php/jpp/article/view/229 |
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Language |
en_US
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Format |
application/pdf
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Publisher |
Proteomics Society, India (PSI)
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