Biophysical characterization of the cetacean morbillivirus haemagglutinin glycoprotein.

"fig 1 design and biophysical characterization of cemv rh-ecto ( a ) molecular evolutionary analysis of the h proteins from members of the morbillivirus genus based on a maximum-likelihood phylogenetic tree and an amino acid sequence identity heatmap both built from a msa of the ncbi protein refseq genbank ids: cdv np_047206 1; cemv np_945029 1; femv yp_009512963 1; gav uqm99546 1; lbbmv-1 ubb42344 1; mbmv ubb97712 1; mev np_056923 1; pdv yp_009177603 1; pomv qwq56142 1; rpv yp_087125 2; romv daz91188 1 srmv yp_133827 2; wnfmv-1 ubb42351 1; the target of this work and orthologs with available protein structure are highlighted as circled in regal-blue (cemv) bright-red (mev) and lilac (cdv) color in the tree (b) cemv genome organization (upper panel) with h gene highlighted in dark gray and structural layout of encoded h protein (lower panel) with tail (champagne) and transmembrane (tacao) domain and stalk (link-water) neck (cornflower) and head (astral) subdomains highlighted in color (c) iupre3 comparative prediction of intrinsic disorder and structural domain organization profile between cemv (regal-blue) cemv rh-ecto (downy cream-brulee astral) cdv (lilac) and mev (bright red) h proteins (d) schematic representation of the structural organization of pcoofy64-sumo3-cemv-rh-ecto construct; his6 -tag and sumo-3 fusion protein are represented as cylinders colored in downy and cream-brulee respectively (e) blue coomassie-stained sds-page (left) and anti-his6 wb analysis (right) of purified cemv rh-ecto (m molecular weight marker) (f) wb coupled to pas glycosylation assay of cemv rh-ecto; magenta-colored bands are indicative of the schiff bonds presence and of the protein glycosylated state (m molecular weight marker; c+ and c- positive and negative controls respectively) (g) nano-dsf analysis of purified cemv rh-ecto; thermal stability and conformational profile is shown by superimposed f330/350 ratio (azure) and first derivative (outrageous-orange); conformational transitions corresponding to inflections in intrinsic fluorescence and related tm 1 (52 0 ?"

"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 the paper."

"Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors."