TY - JOUR
T1 - Biochemical characterization of a GDP-mannose transporter from Chaetomium thermophilum
AU - Premageetha, Gowtham Thambra Rajan
AU - Dhanabalan, Kanaga Vijayan
AU - Bose, Sucharita
AU - Manjunath, Lavanyaa
AU - Joseph, Deepthi
AU - Paz, Aviv
AU - Grandfield, Samuel
AU - Nayak, Vinod
AU - Bredeston, Luis M.
AU - Abramson, Jeff
AU - Ramaswamy, Subramanian
N1 - Funding Information:
RS thanks support from DBT-B-life grant, Grant/Award Number: BT/PR5081/INF/156/2012, DBT-Indo Swedish Grant, Grant/Award Number: BT/IN/SWEDEN/06/SR/2017-18, ESRF Access Program of RCB, Grant/Award Number: BT/INF/22/ SP22660/2017. AP and JA were supported by grant 5R35GM135175-03 from the National Institute of General Medical Sciences. Scientific-Technological Cooperation Program MINCyTArgentina and DST-India (Grant Award Number IN/ 14/09 to LMB and RS). RS thanks support from SERB, India for Grant/Award Number EMR/2016/ 001825. KV thanks support from SERB, India, for a National post-doctoral fellowship. The funders had no role in study design, data collection, analysis, publication decision, or manuscript preparation. The authors declare that they do not have any competing interests.
Publisher Copyright:
Copyright: © 2023 Premageetha et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2023/4
Y1 - 2023/4
N2 - Nucleotide Sugar Transporters (NSTs) belong to the SLC35 family (human solute carrier) of membrane transport proteins and are crucial components of the glycosylation machinery. NSTs are localized in the ER and Golgi apparatus membranes, where they accumulate nucleotide sugars from the cytosol for subsequent polysaccharide biosynthesis. Loss of NST function impacts the glycosylation of cell surface molecules. Mutations in NSTs cause several developmental disorders, immune disorders, and increased susceptibility to infection. Atomic resolution structures of three NSTs have provided a blueprint for a detailed molecular interpretation of their biochemical properties. In this work, we have identified, cloned, and expressed 18 members of the SLC35 family from various eukaryotic organisms in Saccharomyces cerevisiae. Out of 18 clones, we determined Vrg4 from Chaetomium thermophilum (CtVrg4) is a GDP-mannose transporter with an enhanced melting point temperature (Tm) of 56.9̊C, which increases with the addition of substrates, GMP and GDP-mannose. In addition, we report—for the first time—that the CtVrg4 shows an affinity to bind to phosphatidylinositol lipids.
AB - Nucleotide Sugar Transporters (NSTs) belong to the SLC35 family (human solute carrier) of membrane transport proteins and are crucial components of the glycosylation machinery. NSTs are localized in the ER and Golgi apparatus membranes, where they accumulate nucleotide sugars from the cytosol for subsequent polysaccharide biosynthesis. Loss of NST function impacts the glycosylation of cell surface molecules. Mutations in NSTs cause several developmental disorders, immune disorders, and increased susceptibility to infection. Atomic resolution structures of three NSTs have provided a blueprint for a detailed molecular interpretation of their biochemical properties. In this work, we have identified, cloned, and expressed 18 members of the SLC35 family from various eukaryotic organisms in Saccharomyces cerevisiae. Out of 18 clones, we determined Vrg4 from Chaetomium thermophilum (CtVrg4) is a GDP-mannose transporter with an enhanced melting point temperature (Tm) of 56.9̊C, which increases with the addition of substrates, GMP and GDP-mannose. In addition, we report—for the first time—that the CtVrg4 shows an affinity to bind to phosphatidylinositol lipids.
UR - https://www.scopus.com/pages/publications/85153414495
UR - https://www.scopus.com/inward/citedby.url?scp=85153414495&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0280975
DO - 10.1371/journal.pone.0280975
M3 - Article
C2 - 37079572
AN - SCOPUS:85153414495
SN - 1932-6203
VL - 18
JO - PLoS One
JF - PLoS One
IS - 4 April
M1 - e0280975
ER -