Yeast expressed ArtinM shares structure, carbohydrate recognition, and biological effects with native ArtinMInternational Journal of Biological Macromolecules


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ARTICLE IN PRESSG ModelBIOMAC-5398; No. of Pages 9

International Journal of Biological Macromolecules xxx (2015) xxx–xxx

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules j ourna l h o mepa ge: www.elsev ier .com/ locate / i jb iomac

Yeast expressed ArtinM shares structure, carbohy and biological effects with native ArtinM

Nerry Tatiana Cecílioa, Fernanda Caroline Carvalhoa, Yan Liub,

Patrícia Andressa de Almeida Buranelloa, Andre Luiz ZorzettoDouglas d a Soa

Nicholas Ro a Departament ibeirã b Glycosciences m c Department o d Departament e São a r t i c l e i n f o

Article history:

Received 25 February 2015

Received in revised form 28 September 2015

Accepted 29 S

Available onlin




Recombinant p a b s t r a c t

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 1. Introdu

Lectins least one do bohydrates are ubiquit organisms r ∗ Correspon gentes Patogê

Paulo, Avenida

E-mail add ceciliont@gma yan.liu2@impe patburanello@ (A.L. Zorzetto(E.S. Hanna), s njg11@mole.b mcrbarre@fmr http://dx.doi.o 0141-8130/© 0/). this article in press as: N.T. Cecílio, et al., Int. J. Biol. Macromol. (2015), eptember 2015 e xxx latory lectins roteins 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 have a similar subunit formation, oligomerization degree, and carbohydrate recognition specificities that n-ArtinM. We showed that y-ArtinM can exert n-ArtinM biological activities such as erythrocyte agglutination, 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. © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license ( ction are a heterogeneous group of proteins containing at main that selectively and reversibly binds to free caror glycans attached to proteins or lipids [1]. Lectins ous in nature, as they are found in a variety of anging from viruses to humans. The binding of glycans ding author at: Departamento de Biologia Celular e Molecular e Bioanicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São

Bandeirantes 3900, 14049-900 Ribeirão Preto, São Paulo, Brazil. resses: (N.T. Cecílio), (F.C. Carvalho), (Y. Liu), (M. Moncrieffe), (P.A.d.A. Buranello),

Fernandes), (D.D. Luche), (S.G. Soares), (T. Feizi), (N.J. Gay), (M.H.S. Goldman), (M.C.R. Barreira). to the cell surface modifies the plasma membrane and/or promotes signaling that triggers diverse phenomena such as proliferation, migration, and cell death [2]. Due to their important biological activities, lectins are increasingly being studied, and the growing understanding of lectin–carbohydrate interactions at the molecular level provides new and important insights that can be applied in biomedical research.

ArtinM (also known as KM+ or Artocarpin) is a d-mannosebinding lectin that was isolated from the seeds of the Artocarpus heterophyllus (jackfruit). It consists of a 64 kDa homotetramer formed by the association of 16 kDa non-glycosylated subunits [3,4]. This association, although non-covalent, is resistant to detergents such as sodium dodecyl sulfate (SDS), and thermal denaturation is required to obtain electrophoretically detectable monomers from these homotetramers. ArtinM has a high specificity for Man1–3(Man1–6)Man, the trimannoside core of

N-glycans [5]. By interacting with N-glycans of cell surface receptors, ArtinM triggers cellular responses such as neutrophil rg/10.1016/j.ijbiomac.2015.09.062 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license ( Luche , Ebert Seixas Hanna , Sandro Gomes

J. Gayc, Maria Helena S. Goldmand, Maria Cristina o de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de R

Laboratory, Department of Medicine, Imperial College London, London, United Kingdo f Biochemistry, Cambridge University, Cambridge, United Kingdom o de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade ddrate recognition,

Martin Moncrieffec,

Fernandesa, resa, Ten Feizib, que Barreiraa,∗ o Preto, Universidade de São Paulo, Brazil

Paulo, Brazil

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ARTICLE IN PRESSG ModelBIOMAC-5398; No. of Pages 9 2 N.T. Cecílio et al. / International Journal of Biological Macromolecules xxx (2015) xxx–xxx migration and activation by binding to the CXCR2 receptor [6,7], mast cell degranulation by binding to immunoglobulin (Ig) E or its receptor FcR [8,9], and interleukin12 (IL-12) production by inducing activation of macrophages and dendritic cells triggered by interaction