O estudo de albuminas 2S expressas em sementes de Moringa oleifera tem fornecidos evidências de uma grande diversidade de isoformas, em parte, devido à origem multigênica, além do intenso processamento pós-traducional (MPT). Foram consideradas as PTMs: como hidroxilação de prolina, fosforilação de serina ou treonina, oxidação de metionina e cliclização de glutamina N-terminal em piroglutamato (pGlu).
Após alinhamento múltiplo das sequências de aminoácidos deduzidas foi possível agrupá-las em oito grupos distintos. Onde, quatro isoformas foram descritas anteriormente (Mo-CBP3-1, Mo-CBP3-2, Mo-CBP3-3 e Mo-CBP3-4), enquanto que as isoformas Mo-CBP3- 2A (163 resíduos de aminoácidos), Mo-CBP3-2B (162 resíduos), Mo-CBP3-3A (160 resíduos) e Mo-CBP3-3B (160 aa) foram descritas pela primeira vez neste trabalho. Embora existam outras isoformas de albuminas 2S na M. oleifera, as quais não foi possível serem identicadas neste trabalho.
Análises das sequências de aminoácidos, deduzidas de DNA genômico, sugeriram que as isoformas de Mo-CBP3 são sintetizadas como preproproteínas, contendo um peptídeo sinal N-terminal, um propeptídeo N-terminal, uma cadeia menor com cerca de 4 kDa, um peptídeo de ligação entre as duas cadeias, uma cadeia maior com aproximadamente 8 kDa, e uma extensão C-terminal.
Estes resultados constituem importantes avanços na compreensão dos mecanismos pós-traducionais inerentes que operam durante a biossíntese das albuminas 2S nas sementes da M. oleifera.
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APÊNDICE - SUPPLEMENTARY MATERIAL: POST-TRANSLATIONAL MODIFICATIONS REVEAL THAT Mo-CBP3, A 2S MORINGA OLEIFERA CHITIN-
BINDING ALBUMIN, IS A COMPLEX MIXTURE OF ISOFORMS
Table S1. DNA sequences and primer-binding sites of the oligonucleotide primers used in the present work. The sequences of the primers were based on the cDNA sequences, previously determined by Freire et al. (Freire et al., 2015).
Primer Sequence Primer-
binding site Target Mo- CBP3 isoform GenBank accession number of the cDNA sequence P1 P2 5ʹ-CGTCAGTATATCAGAAGCAGTTTAA-3ʹ 5ʹ-AGCTTCGAGCTCTACGAACACACAC-3ʹ 27-51 (+) 670-694 (−) 1/4 KF616830, KF616831 P3 P4 5ʹ-CGTCAGTATATCAGAAGCAGTTTAC-3ʹ 5ʹ-CACGGGGTACATTTGAGCAACTAGC-3ʹ 28-50 (+) 692-716 (−) 2 KF616832 P5 P6 5ʹ-TCAGCAGCAACCAACACCACACCGG-3ʹ 5ʹ-GTTACACCGCTAGTGGCTCTCGTCT-3ʹ 27-51(+) 664-688 (−) 3 KF616833 Fonte: elaborada pelo autor.