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PROTEÓMICA • NÚMERO 1 • FEBRERO 2008

Sede social: Instituto de Biomedicina de Valencia, C.S.I.C.
c/. Jaime Roig, 11. 46010 Valencia
Tel. 96 339 1778 • Fax 96 369 0800
C.I.F.: G-97465629.
Nº. Registro Nacional de Asociaciones: 584180.

http://www.cbm.uam.es/seprot

Junta Directiva:
Juan José Calvete.
Presidente
Instituto de Biomedicina de Valencia
C.S.I.C.
jcalvete@ibv.csic.es
Concha Gil.
Vicepresidenta
Departamento de Microbiología II.
Universidad Complutense de Madrid
conchagil@farm.ucm.es

PROTEÓMICA
Revista de la Sociedad Española
de Proteómica

Jesús V. Jorrín.
Secretario
Universidad de Córdoba
bf1jonoj@uco.es

Número 1, Febrero 2008

David Andreu.
Tesorero
Universidad Pompeu Fabra. Barcelona
david.andreu@upf.edu
Juan Pablo Albar.
Vocal
Centro Nacional de Biotecnología
U.A.M.-C.S.I.C. Madrid
jpalbar@cnb.uam.es
Fernando J. Corrales.
Vocal
Universidad de Navarra
fcorrales@unav.es
Ángela Moreno.
Vocal
Universidad de Córdoba-C.S.I.C.
ge1moloa@uco.es
Jesús M. Vázquez.
Vocal
Centro de Biología Molecular “Severo
Ochoa”. U.A.M.-C.S.I.C. Madrid
jvazquez@cbm.uam.es

COMITÉ EDITORIAL:
Juan J. Calvete (IBV-CSIC, Valencia)
Fernando Corrales (CIMA, Pamplona)
Jesús V. Jorrín (UCO, Córdoba)
Ángela Moreno (CSIC-UCO, Córdoba)
Jesús Vázquez (CBMSO, Madrid)
CORRESPONDENCIA EDITORIAL:
Jesús V. Jorrín Novo. Dpto de Bioquímica y Biología
Molecular, Universidad de Córdoba. Campus de
Rabanales, Ed. Severo Ochoa (C6), 14071 Córdoba.
E-mail: jornadas-proteomica@uco.es.
EDITA:

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PROTEÓMICA • NÚMERO 1 • FEBRERO 2008

PROTEÓMICA. Revista de la Sociedad Española de Proteómica
Editada por: Servicio de Publicaciones de la Universidad de Córdoba
Periodicidad: Semestral (2 números por año: enero-febrero y julio-septiembre).
Contenidos: se publicarán artículos originales, comunicaciones breves, artículos de revisión, artículos de
difusión, opiniones, notas, comentarios sobre cualquier aspecto relacionado con la proteómica. Se priorizarán
artículos originales sobre aspectos metodológicos o de aplicación al estudio de sistemas biológicos. Incluye
información sobre nuestra Sociedad, personas, grupos e instituciones que la componen.
Idioma: será el castellano, aunque se admiten contribuciones en otras lenguas, preferentemente inglés.
Distribución: España y Latinoamérica, aunque pretendemos que tenga un carácter internacional mediante
la distribución a otros países. Se enviarán, sin coste alguno, a los socios de la SEProt, Unidades y Servicios,
así como a instituciones y organizaciones públicas o privadas miembros de la sociedad o con actividad
relevante en el campo de la proteómica.
Publicación: habrá una versión impresa, editada por el Servicio de Publicaciones de la UCO, y una versión
“on-line” que aparecerá en la página web de la SEProt y de la Universidad de Córdoba.
Editores:
Jesús V. Jorrín (UCO, Córdoba)
Juan J. Calvete (IBV-CSIC, Valencia)
Fernando Corrales (CIMA, Pamplona)
Ángela Moreno (CSIC-UCO, Córdoba)
Jesús Vázquez (CBMSO, Madrid)
Correspondencia editorial:
Jesús V. Jorrín Novo. Dpto de Bioquímica y Biología Molecular, Universidad de Córdoba. Campus de Rabanales, Ed. Severo Ochoa (C6), 14071 Córdoba. E-mail: jornadas-proteomica@uco.es.
Instrucciones a los autores:
http://www.cbm.uam.es/seprot/
Envío de los manuscritos:
Mediante correo electrónico a: jornadas-proteomica@uco.es

I.S.S.N.:

1888-0096

Depósito Legal: CO-1005-07
Edita:

Servicio de Publicaciones de la Universidad de Córdoba.
Campus de Rabanales. Ctra. Nacional IV, Km. 396. 14071 CÓRDOBA
Tlfnos.: 957 21 21 65. Fax: 957 21 81 96
http://www.uco.es/publicaciones
Correo electrónico: publicaciones@uco.es

Imprime:

Argos Impresores S.L. Córdoba.

El Servicio de Publicaciones de la Universidad de Córdoba, a los efectos previstos en el artículo 32.1, párrafo 2º, del
vigente TRLPI, se opone expresamente a que cualquiera de las páginas de Proteómica, o partes de ellas, sean utilizadas
para la realización de resúmenes de prensa. Cualquier acto de explotación de sus contenidos precisará de la oportuna
autorización, que será concedida por CEDRO mediante licencia.

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PROTEÓMICA • NÚMERO 1 • FEBRERO 2008

nome (2). Gene recombinant technologies supply agriculture product with great vitality. But the public perception
of genetically modified crops can not be ignored (3).
The safety of transgenic crops should be thoroughly
evaluated based on “substantial equivalence” principle.
The relevant strategies including: substantial equivalent
analysis, toxic tests, protein allergenic study, nutritional assessment, etc. With the development of new technologies,
the approaches of genomic, metabolomics, and proteomics
would be applied to detect the unintended effect.
The present study is focused on the evaluation of the
biochemical changes caused by transformation. Transformed and non-transformed plantlets were compared.
Statistical significant changes, were recorded in levels
of malondialdehyde, other aldehydes, chlorophyll (a, b,
total), phenolics (free and cell wall-linked) and proteins.
Two-dimensional gel electrophoresis and western analysis
has been used to address the in vivo proteomic changes in
transgenic pineapple plants exposed to Finale sublethal
doses harvested after 0, 24, 48, 72 hours and 7 days.
Results indicate that pineapple genetic transformation caused several side effects at the biochemical level
during early stages of plant hardening. These changes
can promote ulterior modifications such as: 1) in the tolerance to stress because levels of malondialdehyde and
other aldehyde were different between transformed and
non-transformed plantlets, 2) in the efficiency of photosynthesis as levels of chlorophyll pigments varied and
3) in the fruit quality as contents of free phenolics were
modified. Proteins, 100µg, were resolved by 2-DE, with
IEF in the 5-8 pH linear range and 12% polyacrylamide
SDS-PAGE. Proteins were visualized by Sypro staining.
The subsequent image comparison and statistic analysis
performed showed changes in protein profiles. Furthermore, about 18 diferential spots in transgenic plants were
analized by matrix-assisted laser desortion ionization
time of flight (MALDI-TOF). showed differential levels
of expression between methods, being some of them absent or unique for a given sample preparation protocol.
At present, pineapple transgenic plants are being studied at the Bioplant Center’s Field Experimental Station
in order to know if market/public concerns regarding genetically modified organisms are scientifically justified or
not. These findings offer interesting perspectives to study
the effect of the herbicide Finale on transgenic pineapple
plants and for pineapple defense programs.

Acknowledgments:
This work was supported by Cuba Ministry of Science, Technology and Environment (CITMA) and Carolina
Fundation, Spain. We thank SCAI (Córdoba University,
Spain) for proteomic studies and analysis .We also want
to thank to Mrs. Julia Martínez and Mrs. Mayda Arzola
for their excellent technical assistance.

References
1.

FAOSTAT (2004) Database, http://appsl.fao.
org

2.

Espinosa P, Lorenzo JC, Iglesias A, Yabor L,
Menéndez E, Borroto J, Hernández L & Arencibia A (2002). Plant Cell Rep. 21: 136-140.

3.

Schubert D (2005). Nat. Biotech. 23: 785-787.

74. Modifications of Arabidopsis thaliana
nuclear proteome in response to
environmental cues
Ribeiro-Pedro, M.1, Ruiz, M. T. 1, Valverde, F. 2, Romero,
J.M. 1
1

Molecular Biology of Starch Metabolism Group, Institute for Plant Biochemistry and Photosynthesis, University of Seville and Spanish Research Council, Seville,
Spain. 2 Plant Development Group, Institute for Plant
Biochemistry and Photosynthesis, Spanish Research
Council and University of Seville, Seville, Spain.
Plant proteomics is far from reaching the standards of
yeast or mammalian systems. Therefore, any new information about plant sub-proteomes has the added value conferred by the scarce knowledge we have and the technical
difficulties involved in data acquisition (Chen and Harmon,
2006). Because plants are sessile organisms, rather than
moving away from changing external conditions like animals do, they respond by modifying their physicochemical
characteristics. This ability confers them a very plastic
physiological behaviour (Casal et al., 2004). For this reason, plant nuclei are packed with molecules that bind to
nucleic acids or are involved in the transfer of incoming and
outgoing signals, making plant nuclear proteome a complex
system to work with (Bae et al., 2003).
In our laboratory we are interested in the signals that
modify gene expression in response to changes in environmental conditions, particularly those dealing with alterations in sugar availability. We have developed a robust
and reproducible protocol to isolate nuclei, extract the
proteome and enrich it in proteins that bind nucleic acids.
It involves chromatographic purification steps, 2DGE and
MS identification of tryptic digests. Using this protocol
on nuclei extract from plants with or without sucrose
addition, we have identified several transcription factors
and other nuclear proteins involved in sugar binding and
signalling that validate our approach. These proteins will
then be analyzed applying other molecular biology and
biochemical tools in an attempt to situate them in the
hierarchical pathway of sugar signalling.

References
Chen, S y Harmon, AC (2007) Advances in plant proteomics. Proteomics 6, 5504-5516.
Casal, JJ, Fankhauser, C, Coupland, G and Blázquez,
MA (2004) Signalling for developmental plasticity. TRENDS in Plant Science 9, 309-314.

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PROTEÓMICA • NÚMERO 1 • FEBRERO 2008

Bae, MS, Cho, EJ, Choi, E-Y and Park, OK (2003)
Analysis of the Arabidopsis nuclear proteome
and its response to cold stress. Plant Journal
36, 652-663.

75. Proteomics analysis of in vitro culture
pineapple (Ananas comosus (L.) Merr.)
shoots
Pérez, A.1, Hernández, M.1, Castillejo, M. A.2, Natalucci,
C.3, Jorrín, J.2*

shoots cultured in vitro. The highest specific protease
activity was recorded in shoots cultured in temporary immersion bioreactors (TIB). Multiplication phase in vitro
did not cause a remarkable protease production in shoots.
Proteome of shoots cultured in different in vitro phase
were compared by 2D- electrophoresis. Molecular mass
of some protein spots were between 21 500- 31 000 Da.
This parameter was similar to those indicated for cysteine
proteases from Bromeliaceae. A protease was detected in
TIB culture media. Retention time in RP-HPLC and molecular mass of the major protein detected in TIB culture
media showed high similarity to stem bromelain.

Laboratorio de Ingeniería Metabólica. Centro de Bioplantas. Universidad de Ciego de Ávila, Ciego de Ávila,
CP 69450, Cuba. TEL: 053-332 224016, Fax: 053-33
266340. 2 Departamento de Bioquímica y Biología Molecular. Universidad de Córdoba. Córdoba. Campus de
Rabanales, Edificio Severo Ochoa (C6), 14071 Córdoba,
España. 3 LIPROVE, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional
de la Plata. La Plata. Argentina. * Corresponding author
(Phone +34957218574; FAX +34957218439; E-mail:
bf1jonoj@uco.es).

Acknowledgments

Bromeliaceae family plants usually contain high concentration of thiol proteases. Pineapple plants (Ananas
comosus) contain several cysteine proteinases (1). In the
literature, these enzymes are called bromelains. Stem
bromelain, in particular, exhibits therapeutic effects: antiinflammatory, digestive, anti-metastasis and anti-tumoral
activities (2, 3). In recent years, it is including in the
drug group modifiers of the biological answer (4). Plant
tissue culture techniques have provided many solutions to
basics questions and practical problems in plant biology.
Therefore, considerable attention has been focused on the
possibility of applying efficient plant tissue culture methods to physiologically active enzymes isolation. In the
present study, we found proteolytic activity in pineapple

References

1

This work was supported by Cuba Ministry of Science, Technology and Environment (CITMA) and Iberoamerican Program of Science and Technology for Development (CYTED), Project IV.22 “Aplicación industrial
de Enzimas Proteolíticas de Vegetales Superiores”, coordined by Dr. Néstor Oscar Caffini. We thank SCAI
(Córdoba University, Spain) for proteomic studies and
analysis. The authors are grateful to Mrs. Carol Carvajal,
Mrs Anabel Pozo and Mrs. Mayelín Mora for their excellent technical assistance.

1.

Rowan, A., Butlle, D. (1994). Methods in Enzimology 244: 555-568.

2.

Tysnes, B., Maurer, H., Porwol, T., Probst, B.,
Bjerkvig, R., Hoover, F. (2001). Neoplasia 3(6):
469-479.

3.

Engwerda, C., Andrew, D., Ladhams, A.,
Mynott, T. (2001). Cellular Immunology 210
(1): 66-75.

4.

Hale, L., Greer, P., Trinh, C., James, C. (2005).
International Immunopharmacology 5: 783-793.

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