caa a22 4500 467894957 CHVBK 20180406152800.0 cr unu---uuuuu 170328e20061101xx s 000 0 eng 10.1007/s11103-006-9038-6 doi (NATIONALLICENCE)springer-10.1007/s11103-006-9038-6 Molecular cloning, expression, and cytokinin (6-benzylaminopurine) antagonist activity of peanut ( Arachis hypogaea ) lectin SL-I [Elektronische Daten] [Monika Pathak, Bharat Singh, Amit Sharma, Praveen Agrawal, Santosh Pasha, Hasi Das, Rakha Das] Isolation and purification of a α-methyl-mannoside specific lectin (SL-I) of peanut was reported earlier [Singh and Das (1994) Glycoconj J 11:282-285]. Native SL-I is a glycoprotein having ∼31kDa subunit molecular mass and forms dimer. The gene encoding this lectin is identified from a 6-day old peanut root cDNA library by anti-SL-I antibody and N-terminal amino acid sequence homology to the native lectin. Nucleotide sequence derived amino acid sequence of the re-SL-I shows amino acid sequence homology with the N-terminal and tryptic digests' amino acid sequence of the native SL-I (nSL-I). Presence of a putative glycosylation (QNPS) site and a hydrophobic adenine-binding (VLVSYDANS) site is also identified in SL-I. Homology modeling of the lectin suggests it to be an archetype of legume lectins. It is expressed as a ~30kDa apoprotein in E. coli and has the carbohydrate specificity and secondary structure identical to its natural counterpart. The lectin SL-I inhibits cytokinin 6-benzylaminopurine (BA)-induced "delayed leaf senescence” and "cotyledon expansion”. Equilibrium dialysis revealed a single high-affinity binding site for adenine (7.6×10−6M) and BA (1.09×10−5M) in the SL-I dimer and thus suggesting that the cytokinin antagonist effect of SL-I is mediated by the direct interaction of SL-I with BA. Springer Science+Business Media B.V., 2006 Peanut lectin SL-I nationallicence Gene expression nationallicence Cytokinin 6-benzylaminopurine nationallicence Cytokinin antagonist activity nationallicence PBS : phosphate-buffered saline nationallicence nSL-I/SL-I : native peanut stem lectin nationallicence re-SL-I : recombinant SL-I nationallicence Con A : Concanavalin A nationallicence LBL : lima bean lectin nationallicence WBA I and WBA II : winged bean basic- and acidic- agglutinin respectively nationallicence PHA-E : Phaseolus vulgaris erythroagglutinin nationallicence PAL : Pterocarpus angolensis man/glc specific seed lectin nationallicence SBA : soybean lectin nationallicence ANS : 1,8-anilino-1-naphthalenesulfonic acid nationallicence BA : 6-benzylaminopurine nationallicence PMSF : phenylmethylsulfonylfluoride nationallicence Pathak Monika Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Singh Bharat Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Sharma Amit Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Agrawal Praveen Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Pasha Santosh Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Das Hasi Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Das Rakha Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut Plant Molecular Biology Kluwer Academic Publishers 62/4-5(2006-11-01), 529-545 0167-4412 62:4-5<529 2006 62 11103 https://doi.org/10.1007/s11103-006-9038-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-9038-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Pathak Monika Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Singh Bharat Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Sharma Amit Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Agrawal Praveen Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Pasha Santosh Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Das Hasi Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 700 1- Das Rakha Unit of Proteomics and Comparative Genomics, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, 110 007, Delhi, India aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 62/4-5(2006-11-01), 529-545 0167-4412 62:4-5<529 2006 62 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer