Combining multidimensional liquid chromatography and MALDI-TOF-MS for the fingerprint analysis of secreted peptides from the unexplored sea anemone species Phymanthus crucifer

Autor(es)

Rodriguez, Armando A. ^[1] ; Staendker, Ludger ^[2] ; Zaharenko, Andre J. ^[3] ; Garateix, Anoland G. ^[1] ; Forssmann, Wolf-Georg ^[2] ; Beress, Laszlo ^[2] ; Valdes, Olga ^[1] ; Hernandez, Yasnay ^[1] ; Laguna, Abilio ^[1]

Afiliação do(s) autor(es)

^[1] Ctr Bioprod Marinos CEBIMAR, Havana - Cuba

^[2] Hannover Med Sch, D-30625 Hannover - Germany

^[3] Inst Butantan, Genet Lab, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES; v. 903, p. 30-39, AUG 15 2012.
• Citações Web of Science: 9

Resumo

Sea anemones are sources of biologically active proteins and peptides. However, up to date few peptidomic studies of these organisms are known; therefore most species and their peptide diversity remain unexplored. Contrasting to previous venom peptidomic works on sea anemones and other venomous animals, in the present study we combined pH gradient ion-exchange chromatography with gel filtration and reversed-phase chromatography, allowing the separation of the 1-10 kDa polypeptides from the secretion of the unexplored sea anemone Phymanthus crucifer (Cnidaria/Phymanthidae). This multidimensional chromatographic approach followed by MALDI-TOF-MS detection generated a peptide fingerprint comprising 504 different molecular mass values from acidic and basic peptides, being the largest number estimated for a sea anemone exudate. The peptide population within the 2.0-3.5 kDa mass range showed the highest frequency whereas the main biomarkers comprised acidic and basic peptides with molecular masses within 2.5-6.9 kDa, in contrast to the homogeneous group of 4-5 kDa biomarkers found in sea anemones such as B. granulifera and B. cangicum (Cnidaria/Actiniidae). Our study shows that sea anemone peptide fingerprinting can be greatly improved by including pH gradient ion-exchange chromatography into the multidimensional separation approach, complemented by MALDI-TOF-MS detection. This strategy allowed us to find the most abundant and unprecedented diversity of secreted components from a sea anemone exudate, indicating that the search for novel biologically active peptides from these organisms has much greater potential than previously predicted. (C) 2012 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 07/56525-3 - Construção de uma biblioteca de cDNA dos tentáculos da anêmona do mar Bunodosoma cangicum e clonagem de neurotoxinas
• Beneficiário: Andre Junqueira Zaharenko
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado