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Recombinant pre-pro-Concanavalin A (jack bean) is stable but of low solubility

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Abstract

The cDNA for pre-pro-Concanavalin A (pre-pro-ConA) was cloned into the cytoplasmic expression vector pKK233-2 to give rise to pCONEXP2 which was used to express the lectin precursor. Pre-pro-ConA is stable and is not transposed and ligated to form the mature protein. No signal peptide removal is observed. The solubility of pre-pro-ConA could not be increased by guanidine hydrochloride denaturation/dilution treatment.

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Abbreviations

ConA:

Concanavalin A

IPTG:

isopropyl-Β-D-thiogalactopyranoside

PMSF:

phenyl methyl sulphonyl flouride

References

  • Amann E and Brosius J 1985 ‘ATG’ vectors for regulated expression of cloned genes inEscherichia coli;Gene 40 183–190

    Article  CAS  Google Scholar 

  • Bowles D J, Marcus S E, Pappin D J C, Findlay J B C, Eliopoulos E, Maycox P R and Burgess J 1986 Post-translational processing of ConA precursors in Jack Bean cotyledons;J. Cell Biol. 102 1284–1297

    Article  CAS  Google Scholar 

  • Carrington D M, Aufret A and Hanke D E 1985 Polypeptide ligation occurs during post-translational processing of Concanavalin A;Nature (London) 313 64–67

    Article  CAS  Google Scholar 

  • Cunningham B A, Wang J L, Waxdal M J and Edelman G M 1975 The covalent and the three dimensional structure of Concanavalin A;J. Biol. Chem. 250 1503–1512

    CAS  PubMed  Google Scholar 

  • Gatenby A A, Boccara M, Baulcombe D C and Rothstein S J 1986 Expression of wheat α-amylase inEscherichia coli;Gene 45 11–18

    Article  CAS  Google Scholar 

  • Goldstein I J and Poretz R D 1986 Isolation and characterization and carbohydrate specificity of lectins; inThe lectins: Properties, functions and applications in biology and medicine (eds) I E Liener, N Sharon and I J Goldstein (London: Academic Press) pp

    Google Scholar 

  • Harlow E and Lane D 1989Antibodies: A laboratory manual (New York: Cold Spring Laboratory)

    Google Scholar 

  • Hemperly J J and Cunningham B A 1985 Circular permutation of amino acid sequences among legume lectins;Trends Biochem. Sci. 8 100–102

    Article  Google Scholar 

  • Herman E M, Shannon L M and Chrispeels M J 1986 Concanavalin A is synthesized as a glycosylated precursor;Planta 165 23–29

    Article  Google Scholar 

  • Jones D H, Min W, Dincturk H B, Dunn A J, Williams R P and Li M 1996 Concanavalin A and precursors used to investigate its extraordinary post-translational processing; inLectins: Biology, biochemistry, clinical biochemistry (eds) E Van Driessche, P Rouge, S Beeckmans and T C Bog-Hansen (Denmark: Textop) vol. 11, pp 70–73

    Google Scholar 

  • Kozak M 1983 Comparison and initiation of protein synthesis in prokaryotes, eukaryotes and organelles;Microbiol. Rev. 47 1–45

    CAS  PubMed  PubMed Central  Google Scholar 

  • Min W, Dunn A J and Jones D H 1992 Non-glycosylated recombinant pro-Concanavalin A is active without polypeptide cleavage;EMBO J. 11 1303–1307

    Article  CAS  Google Scholar 

  • Min W and Jones D H 1992 Stability and detection of recombinant pre-pro-Concanavalin A after cytoplasmic expression inEscherichia coli;FEBS Lett. 301 315–318

    Article  CAS  Google Scholar 

  • Min W and Jones D H 1994In vitro splicing of Concanavalin A is catalysed by asparaginyl endopeptidase;Nature Struct. Biol. 1 502–504

    Article  CAS  Google Scholar 

  • Mitraki A and King J 1990 Protein folding intermediates and inclusion body formation;Bio/Technology 7 690–697

    Google Scholar 

  • Sambrook J, Fritsch E F and Maniatis T 1989Molecular cloning, Second edition (New York: Cold Spring Harbour Laboratory)

    Google Scholar 

  • Stubbs M E, Carver J P and Dunn R J 1986 Production of pea lectin inEscherichia coli;J. Biol. Chem. 261 6141–6144

    CAS  PubMed  Google Scholar 

  • Williams T J, Shafer J A and Goldstein I J 1978 Heterogeneity of Concanavalin A as detected by its binding to p-nitrophenyl 2-0-α-D-mannopyronoside;J. Biol. Chem. 253 8533–8537

    CAS  PubMed  Google Scholar 

  • Yamauchi D and Minimikawa T 1990 Structure of the gene coding for Concanavalin A fromCanavalia gladiata and its expression inEscherichia coli cells;FEBS Lett. 260 127–130

    Article  CAS  Google Scholar 

  • Yamauchi D, Nakamura K, Asahi T, Minimikawa T 1988 cDNAs for canavalin and Concanavalin A fromCanavalia gladiata seeds. Nucleotide sequence of cDNA and canavalin and RNA blot analysis of canavalin and Concanavalin A in developing seeds;Eur. J. Biochem. 170 515–520

    Article  CAS  Google Scholar 

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Author notes

  1. H. Benan Dincturk

    Present address: Faculty of Sciences and Letters, Department of Molecular Biology and Genetics, Istanbul Technical University, Maslak, 80626, Istanbul, Turkey

Authors and Affiliations

  1. Molecular Biology Research Group, School of Biological Sciences, University of Wales Swansea, Singleton Park, SA2 8PP, Swansea, UK

    H. Benan Dincturk, Anthony J. Dunn & D. Hugh Jones

Authors
  1. H. Benan Dincturk
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  2. Anthony J. Dunn
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  3. D. Hugh Jones
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Correspondence to H. Benan Dincturk.

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Dincturk, H.B., Dunn, A.J. & Jones, D.H. Recombinant pre-pro-Concanavalin A (jack bean) is stable but of low solubility. J. Biosci. 26, 635–640 (2001). https://doi.org/10.1007/BF02704761

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  • DOI: https://doi.org/10.1007/BF02704761

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