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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
 
Creator Rani, Priya
Gautam, Gunjan
Rao, Kongara Hanumantha
Ghosh, Swagata
Gourinath, Samudrala
Dhar, Suman Kumar
Datta, Asis
 
Subject N-acetylglucosamine (GlcNAc)
GlcNAc Inducible Gene 2 (GIG2)
DUF (Domains of Unknown Function) family of proteins
Crystallography
 
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.
 
Date 2018-06-25T08:13:11Z
2018-06-25T08:13:11Z
2017
 
Type Article
 
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
 
Language en_US
 
Format application/pdf
 
Publisher Proteomics Society, India (PSI)