Yeast expressed ArtinM shares structure, carbohydrate recognition, and biological effects with native ArtinM

Abstract

Recent advances in glycobiology have revealed the essential role of lectins in deciphering the glycocodes at the cell surface to generate important biological signaling responses. ArtinM, a d-mannose-binding lectin isolated from the seeds of jackfruit (Artocarpus heterophyllus), is composed of 16 kDa subunits that are associated to form a homotetramer. Native ArtinM (n-ArtinM) exerts immunomodulatory and regenerative effects, but the potential pharmaceutical applicability of the lectin is highly limited by the fact that its production is expensive, laborious, and impossible to be scaled up. This led us to characterize a recombinant form of the lectin obtained by expression in Saccharomyces cerevisiae (y-ArtinM). In the present study, we demonstrated that y-ArtinM is similar to n-ArtinM in subunit arrangement, oligomerization and carbohydrate binding specificity. We showed that y-ArtinM can exert n-ArtinM biological activities such as erythrocyte agglutination, stimulation of neutrophil migration and degranulation, mast cell degranulation, and induction of interleukin-12 and interleukin-10 production by macrophages. In summary, the expression of ArtinM in yeast resulted in successful production of an active, recombinant form of ArtinM that is potentially useful for pharmaceutical application.

CD14 is critical for TLR2-mediated M1 macrophage activation triggered by N-glycan recognition

“…Although ArtinM recognizes primarily Manα1-3(Manα1-6)Man, which is the common core of N-glycans, the fine specificity of its binding is further determined by a secondary subsite of recognition. It interacts with other carbohydrate residues (Fuc and GlucNAc), which may be contained in the N-glycan, resulting in the particular binding property exhibited by this lectin 32 , 33 , 54 .…”

“…This activity was attributed to ArtinM interaction with TLR2 N-glycans 31 , which is followed by IL-12 production and establishment of Th1 immunity. The ArtinM effects were shown to depend on carbohydrate recognition because direct binding to TLR2 was inhibited by the trimannoside specifically targeted by ArtinM 32 , 33 , which constitutes the core of N-glycans. These data demonstrated that a carbohydrate recognition protein might function as a TLR agonist and exert immunomodulatory activity.…”

ArtinM Mediates Murine T Cell Activation and Induces Cell Death in Jurkat Human Leukemic T Cells

“…This research confirmed the preference of the ArtinM primary site for probes having the core structure Manα1–3[Manα1–6]Manβ1–4. It also showed the contribution of the secondary site of ArtinM to enhancing the affinity of the CRD for probes containing Fucα1–6 or GlcNAcα1–2 associated with Manα1–3[Manα1–6]Manβ1–4 [ 47 , 48 , 49 ]. The core structure Manα1–3[Manα1–6]Manβ1–4 recognized by ArtinM is also targeted by Morniga M [ 50 , 51 ], which is consistent with the marked correlation between the glycan-binding specificities and phylogenies of ArtinM and Morniga M [ 46 ].…”

Glycan microarray analysis of the carbohydrate-recognition specificity of native and recombinant forms of the lectin ArtinM

“…n-ArtinM was obtained from a saline extract of Artocarpus heterophyllus (jackfruit) seeds [6] . Saccharomyces cerevisiae BJ3501 was used to express y-ArtinM and the lectin was obtained by yeast lysis [1] . n-ArtinM and y-ArtinM were purified by affinity chromatography on a D -mannose column coupled to AKTA Purifier (GE Healthcare, Bio-Science Inc. Germany), previously equilibrated with phosphate-buffered saline (PBS) containing 0.5 M NaCl.…”

“… This article contains data related to the researc.h article entitled “Yeast-derived ArtinM shares structure, carbohydrate recognition, and biological effects with native ArtinM” by Cecílio et al (2015) [1] . ArtinM, a D -mannose-binding lectin isolated from the seeds of Artocarpus heterophyllus, exerts immunomodulatory and regenerative activities through its Carbohydrate Recognition Domain (CRD) (Souza et al, 2013; Mariano et al, 2014 [2] , [3] ).…”