PDBsum entry 6plh

Immune system

PDB id: 6plh

Contents

Protein chains

219 a.a.

219 a.a.

Ligands

THR-TRP-SER-GLU-TRP-SER-ASP-PRO

PO4

MAN

Waters ×370

PDB id:

6plh

Name:

Immune system

Title:

Fab fragment complexed with c-mannosylated tryptophan peptide

Structure:

Fab 5g12 light chain. Chain: a. Fab 5g12 heavy chain. Chain: b. Interleukin-21 receptor. Chain: c. Fragment: unp residues 202-232. Synonym: il-21r,novel interleukin receptor. Engineered: yes

Source:

Mus musculus. Organism_taxid: 10090. Homo sapiens. Human. Organism_taxid: 9606. Gene: il21r, nilr, unq3121/pro10273. Expressed in: komagataella pastoris. Expression_system_taxid: 4922

Resolution:

1.60Å R-factor: 0.191 R-free: 0.221

Authors:

A.John,M.A.Jarva,E.D.Goddard-Borger

Key ref:

A.John et al. (2021). Yeast- and antibody-based tools for studying tryptophan C-mannosylation. Nat Chem Biol, 17, 428-437. PubMed id: 33542533 DOI: 10.1038/s41589-020-00727-w

Date:

30-Jun-19 Release date: 08-Jul-20

Protein chain

No UniProt id for this chain

Struc: 219 a.a.

Protein chain

No UniProt id for this chain

Struc: 219 a.a.

DOI no: 10.1038/s41589-020-00727-w

Nat Chem Biol 17:428-437 (2021)

PubMed id: 33542533

Yeast- and antibody-based tools for studying tryptophan C-mannosylation.

A.John, M.A.Järvå, S.Shah, R.Mao, S.Chappaz, R.W.Birkinshaw, P.E.Czabotar, A.W.Lo, N.E.Scott, E.D.Goddard-Borger.

ABSTRACT

Tryptophan C-mannosylation is an unusual co-translational protein modification performed by metazoans and apicomplexan protists. The prevalence and biological functions of this modification are poorly understood, with progress in the field hampered by a dearth of convenient tools for installing and detecting the modification. Here, we engineer a yeast system to produce a diverse array of proteins with and without tryptophan C-mannosylation and interrogate the modification's influence on protein stability and function. This system also enabled mutagenesis studies to identify residues of the glycosyltransferase and its protein substrates that are crucial for catalysis. The collection of modified proteins accrued during this work facilitated the generation and thorough characterization of monoclonal antibodies against tryptophan C-mannosylation. These antibodies empowered proteomic analyses of the brain C-glycome by enriching for peptides possessing tryptophan C-mannosylation. This study revealed many new modification sites on proteins throughout the secretory pathway with both conventional and non-canonical consensus sequences.