A conserved asparagine residue in the inner surface of BRI1 superhelix is essential for protein native conformation.

Asparagine-linked glycosylation (ALG, N-glycosylation) is one of the most prevalent protein modifications in eukaryotes and regulates protein folding, trafficking and function. Recently, we reported that the mutation of N154Q significantly led to the ER retention of brassinosteroids insensitive 1 (BRI1), the receptor of brassinosteroids (BRs). However, the mechanism of how the N154 site affects BRI1 structure is still not completely clear. In current study, we found that the removal of N ¹⁵⁴ -glycan with S156A replacement significantly enhanced the ability of bri1 to complement bri1-301 mutant and plasma membrane localization compared with N154Q. In addition, the various mutations on N154 site resulted in bri1 retention in the ER, except for N154D. The 3D modeling suggested that there existed polar contacts around N154 site and the mutations not only destroyed the addition of N-glycan on the site, but also led to the disorder of hydrogen bonds formation. The sequence analysis showed that the N275 shared more similarity with N154 site and the removal of N ²⁷⁵ -glycan further enhanced the retention of bri1 carrying S156A mutation in the ER. Our results showed that N154 was special and essential for maintaining BRI1 structure and explored the role of those residues and key N-glycans lying in the LRR inner surface on protein conformation.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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