TY - JOUR
T1 - Agl28 and Agl29 are key components of a Halobacterium salinarum N-glycosylation pathway
AU - Vershinin, Zlata
AU - Zaretsky, Marianna
AU - Guan, Ziqiang
AU - Eichler, Jerry
N1 - Publisher Copyright:
© 2023 The Author(s). Published by Oxford University Press on behalf of FEMS.
PY - 2023/3/2
Y1 - 2023/3/2
N2 - Although Halobacterim salinarum provided the first example of N-glycosylation outside the Eukarya, only recently has attention focused on delineating the pathway responsible for the assembly of the N-linked tetrasaccharide decorating selected proteins in this haloarchaeon. In the present report, the roles of VNG1053G and VNG1054G, two proteins encoded by genes clustered together with a set of genes demonstrated to encode N-glycosylation pathway components, were considered. Relying on both bioinformatics and gene deletion and subsequent mass spectrometry analysis of known N-glycosylated proteins, VNG1053G was determined to be the glycosyltransferase responsible for addition of the linking glucose, while VNG1054G was deemed to be the flippase that translocates the lipid-bound tetrasaccharide across the plasma membrane to face the cell exterior, or to contribute to such activity. As observed with Hbt. salinarum lacking other components of the N-glycosylation machinery, both cell growth and motility were compromised in the absence of VNG1053G or VNG1054G. Thus, given their demonstrated roles in Hbt. salinarum N-glycosylation, VNG1053G and VNG1054G were re-annotated as Agl28 and Agl29, according to the nomenclature used to define archaeal N-glycosylation pathway components.
AB - Although Halobacterim salinarum provided the first example of N-glycosylation outside the Eukarya, only recently has attention focused on delineating the pathway responsible for the assembly of the N-linked tetrasaccharide decorating selected proteins in this haloarchaeon. In the present report, the roles of VNG1053G and VNG1054G, two proteins encoded by genes clustered together with a set of genes demonstrated to encode N-glycosylation pathway components, were considered. Relying on both bioinformatics and gene deletion and subsequent mass spectrometry analysis of known N-glycosylated proteins, VNG1053G was determined to be the glycosyltransferase responsible for addition of the linking glucose, while VNG1054G was deemed to be the flippase that translocates the lipid-bound tetrasaccharide across the plasma membrane to face the cell exterior, or to contribute to such activity. As observed with Hbt. salinarum lacking other components of the N-glycosylation machinery, both cell growth and motility were compromised in the absence of VNG1053G or VNG1054G. Thus, given their demonstrated roles in Hbt. salinarum N-glycosylation, VNG1053G and VNG1054G were re-annotated as Agl28 and Agl29, according to the nomenclature used to define archaeal N-glycosylation pathway components.
KW - Halobacterium salinarum
KW - N-glycosylation
KW - archaea
KW - glycoproteins
KW - glycosyltransferase
UR - http://www.scopus.com/inward/record.url?scp=85150666528&partnerID=8YFLogxK
U2 - 10.1093/femsle/fnad017
DO - 10.1093/femsle/fnad017
M3 - Article
C2 - 36866517
AN - SCOPUS:85150666528
SN - 0378-1097
VL - 370
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
M1 - fnad017
ER -