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Purification and Characterization of a Lectin with High Hemagglutination Property Isolated from Allium altaicum

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Abstract

A lectin was purified from the leaves of Allium altaicum and corresponding gene was cloned. The lectin namely Allium altaicum agglutinin (AAA) was ~24 kDa homodimeric protein and similar to a typical garlic leaf lectin. It was synthesized as 177 amino acid residues pre-proprotein, which consisted of 28 and 43 amino acid long N and C-terminal signal peptides, respectively. The plant expressed this protein more in scapes and flowers in comparison to the bulbs and leaves. Hemagglutination activity (with rabbit erythrocytes) was 1,428 fold higher as compared to Allium sativum leaf agglutinin (ASAL) although, the insecticidal activity against cotton aphid (Aphis gossypii) was relatively low. Glycan array revealed that AAA had higher affinity towards GlcAb1-3Galb as compared to ASAL. Homology analysis showed 57–94% similarity with other Allium lectins. The mature protein was expressed in E. coli as a fusion with SUMO peptide in soluble and biologically active form. Recombinant protein retained high hemagglutination activity.

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Abbreviations

AAA:

Allium altaicum agglutinin

ASAL:

Allium sativum leaf agglutinin

EDTA:

Ethylenediamine tetra acetic acid

SUMO:

Small ubiquitin like modifier

RFU:

Relative fluorescence units

GNA:

Galanthus nivalis agglutinin

RT-PCR:

Reverse transcriptase PCR

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Acknowledgments

This work has been funded by Council of Scientific and Industrial Research (CSIR), Government of India, Supra Institutional Projects (SIP005). SKU, SS, RS and NKD acknowledge CSIR and PR acknowledge Indian Council of Medical Research, Government of India for respective Senior Research Fellowship. RT is thankful to DST for JC bose fellowship. The authors are also thankful to Moti Lal, SMH Abidi, Rajesh Srivastava and Aquila Bano for technical support.

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Authors and Affiliations

  1. Council of Scientific and Industrial Research, National Botanical Research Institute, Rana Pratap Marg, Lucknow, Uttar Pradesh, 226001, India

    Santosh Kumar Upadhyay, Sharad Saurabh, Rahul Singh, Preeti Rai, Neeraj Kumar Dubey, K. Chandrashekar & P. K. Singh

  2. Department of Biotechnology, National Agri-Food Biotechnology Institute, C-127, Phase VIII, Industrial Area, SAS Nagar, Mohali, Punjab, 160071, India

    Rakesh Tuli

  3. National Bureau of Plant Genetic Resources, Regional Station, Bhowali, Nainital, Uttarakhand, 263132, India

    Kuldeep Singh Negi

Authors
  1. Santosh Kumar Upadhyay
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  2. Sharad Saurabh
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  3. Rahul Singh
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  5. Neeraj Kumar Dubey
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  7. Kuldeep Singh Negi
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  8. Rakesh Tuli
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  9. P. K. Singh
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Corresponding author

Correspondence to P. K. Singh.

Additional information

Santosh Kumar Upadhyay and Sharad Saurabh Contributed equally to the results of this study.

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Upadhyay, S.K., Saurabh, S., Singh, R. et al. Purification and Characterization of a Lectin with High Hemagglutination Property Isolated from Allium altaicum . Protein J 30, 374–383 (2011). https://doi.org/10.1007/s10930-011-9342-0

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