Cystine-mediated oligomerization of the Atlantic salmon serum C-type lectin

Hudson, David M.; Mattatall, Neil R.; Uribe, Elke; Richards, Robert C.; Gong, Huansheng; Ewart, K. Vanya

doi:10.1016/j.bbapap.2010.11.004

Cited by 6 publications

(6 citation statements)

References 44 publications

(59 reference statements)

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“…The double-loop structure of the CRD is stabilized by disulphide bonds formed by four Cys residues conserved in all the C-type lectins [2,4]. In addition, C-type lectin oligomerization, that is mediated by disulphide bonds too, is probably playing an important role in their function and selectivity [22]. It is very likely that the additional Cys residue present in BLL2 is responsible of the higher activity of this protein when compared to BLL1.…”

Section: Discussionmentioning

confidence: 99%

High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from <em>Spodoptera exigua</em> (Lepidoptera: Noctuidae)

Gasmi

Ferré

Herrero

2017

Preprint

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Abstract:Lectins are carbohydrate-interacting proteins playing a pivotal role in multiple physiological and developmental aspects of all organisms. They can specifically interact with different bacterial and viral pathogens through the carbohydrate-recognition domains (CRD). In addition, lectins are also of biotechnological interest because of their potential use as biosensor for capturing and identification of bacterial species. In this work, we have characterized the bacterial agglutination properties of three C-type lectins from the Lepidoptera Spodoptera exigua. One of these lectins, BLL2, was able to agglutinate cells from a broad range of bacterial species at an extremely low concentration, becoming a very interesting protein to be used as biosensor or other biotechnological applications involving bacterial capturing.

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Section: Discussionmentioning

confidence: 99%

High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from <em>Spodoptera exigua</em> (Lepidoptera: Noctuidae)

Gasmi

Ferré

Herrero

2017

Preprint

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“…The dried sample was dissolved in 30 lL ddH 2 O and the oligomeric organization of rSSL was analyzed by non-reducing SDS-PAGE on a 12.5 % polyacrylamide gel (NEXT GEL, Amresco) followed by western blotting on a PVDF membrane. Recombinant SSL expressed in E. coli as previously described (Hudson et al 2011) was analyzed under the same conditions.…”

Section: Oligomerization Analysismentioning

confidence: 99%

“…The luminescent signal emitted by the enhanced chemiluminescence substrate (Plus-ECL, PerkinElmer) was detected with X-ray film. Standards used to estimate rSSL concentration in western blot analyses were 30-500 ng pure native SSL (E. Uribe et al, manuscript submitted) and 30-500 ng pure rSSL from E. coli (Hudson et al 2011). …”

Section: Sds-page and Western Blot Analysismentioning

confidence: 99%

“…A computer-generated model of the SSL-2 isoform subunit showed the three expected intramolecular disulfides and suggested that the two remaining cysteines are moderately exposed to the solvent. Both of these exposed cysteines were found to contribute to oligomerization of SSL (Hudson et al 2011). …”

Section: Introductionmentioning

confidence: 99%

“…However, SSL is a cystine-rich, oligomeric protein that has proven to be a challenging target for recombinant protein expression. Expression of soluble rSSL was achieved in Escherichia coli (Hudson et al 2011), but the yield was very low and variable.…”

Section: Introductionmentioning

confidence: 99%

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Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

et al. 2012

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The Atlantic salmon (Salmo salar) serum lectin (SSL) is a soluble C-type lectin that binds bacteria, including salmon pathogens. This lectin is a cysteine-rich oligomeric protein. Consequently, a Drosophila melanogaster expression system was evaluated for use in expressing SSL. A cDNA encoding SSL was cloned into a vector designed to express it as a fusion protein with a hexahistidine tag, under the control of the Drosophila methallothionein promoter. The resulting construct was stably transfected into Drosophila S2 cells. After CdCl 2 induction, transfected S2 cells secreted recombinant SSL into the cell culture medium. A cell line derived from stably transformed polyclonal cell populations expressing SSL was used for large-scale expression of SSL. Recombinant SSL was purified from the culture medium using a two-step purification scheme involving affinity binding to yeast cells and metal-affinity chromatography. Although yields of SSL were very low, correct folding and functionality of the recombinant SSL purified in this manner was demonstrated by its ability to bind to Aeromonas salmonicida. Therefore, Drosophila S2 cells may be an ideal system for the production of SSL if yields can be increased.

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A new mucin-binding lectin from the marine sponge Aplysina fulva (AFL) exhibits antibiofilm effects

Carneiro

Viana

Torres

et al. 2019

Archives of Biochemistry and Biophysics

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Cystine-mediated oligomerization of the Atlantic salmon serum C-type lectin

Cited by 6 publications

References 44 publications

High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from <em>Spodoptera exigua</em> (Lepidoptera: Noctuidae)

High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from <em>Spodoptera exigua</em> (Lepidoptera: Noctuidae)

Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

A new mucin-binding lectin from the marine sponge Aplysina fulva (AFL) exhibits antibiofilm effects

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