An extracellular transglutaminase is required for apple pollen tube growth

Alessia Di Sandro, Stefano Del Duca, Elisabetta Verderio, Alan J. Hargreaves, Alessandra Scarpellini, Giampiero Cai, Mauro Cresti, Claudia Faleri, Rosa A. Iorio, Shigehisa Hirose, Yutaka Furutani, Ian G.C. Coutts, Martin Griffin, Philip L.R. Bonner, Donatella Serafini-Fracassini

Research output: Contribution to journalArticle

Abstract

An extracellular form of the calcium-dependent protein-cross-linking enzyme TGase (transglutaminase) was demonstrated to be involved in the apical growth of Malus domestica pollen tube. Apple pollen TGase and its substrates were co-localized within aggregates on the pollen tube surface, as determined by indirect immunofluorescence staining and the in situ cross-linking of fluorescently labelled substrates. TGase-specific inhibitors and an anti-TGase monoclonal antibody blocked pollen tube growth, whereas incorporation of a recombinant fluorescent mammalian TGase substrate (histidine-tagged green fluorescent protein: His6-Xpr-GFP) into the growing tube wall enhanced tube length and germination, consistent with a role of TGase as a modulator of cell wall building and strengthening. The secreted pollen TGase catalysed the cross-linking of both PAs (polyamines) into proteins (released by the pollen tube) and His6-Xpr-GFP into endogenous or exogenously added substrates. A similar distribution of TGase activity was observed in planta on pollen tubes germinating inside the style, consistent with a possible additional role for TGase in the interaction between the pollen tube and the style during fertilization.
LanguageEnglish
Pages261-271
Number of pages11
JournalBiochemical Journal
Volume429
Issue number2
DOIs
Publication statusPublished - 5 May 2010

Fingerprint

Pollen Tube
Transglutaminases
Malus
Growth
Substrates
Pollen
Polyamines
Indirect Fluorescent Antibody Technique
Germination
Green Fluorescent Proteins
Histidine
Fertilization
Cell Wall
Modulators
Proteins
Monoclonal Antibodies
Cells
Staining and Labeling
Calcium

Keywords

  • extracellular space
  • germination
  • green fluorescent proteins
  • immunohistochemistry
  • malus
  • biological models
  • genetically modified plants
  • pollen tube
  • recombinant proteins
  • substrate specificity
  • transglutaminases

Cite this

Di Sandro, A., Del Duca, S., Verderio, E., Hargreaves, A. J., Scarpellini, A., Cai, G., ... Serafini-Fracassini, D. (2010). An extracellular transglutaminase is required for apple pollen tube growth. Biochemical Journal, 429(2), 261-271. https://doi.org/10.1042/BJ20100291
Di Sandro, Alessia ; Del Duca, Stefano ; Verderio, Elisabetta ; Hargreaves, Alan J. ; Scarpellini, Alessandra ; Cai, Giampiero ; Cresti, Mauro ; Faleri, Claudia ; Iorio, Rosa A. ; Hirose, Shigehisa ; Furutani, Yutaka ; Coutts, Ian G.C. ; Griffin, Martin ; Bonner, Philip L.R. ; Serafini-Fracassini, Donatella. / An extracellular transglutaminase is required for apple pollen tube growth. In: Biochemical Journal. 2010 ; Vol. 429, No. 2. pp. 261-271.
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abstract = "An extracellular form of the calcium-dependent protein-cross-linking enzyme TGase (transglutaminase) was demonstrated to be involved in the apical growth of Malus domestica pollen tube. Apple pollen TGase and its substrates were co-localized within aggregates on the pollen tube surface, as determined by indirect immunofluorescence staining and the in situ cross-linking of fluorescently labelled substrates. TGase-specific inhibitors and an anti-TGase monoclonal antibody blocked pollen tube growth, whereas incorporation of a recombinant fluorescent mammalian TGase substrate (histidine-tagged green fluorescent protein: His6-Xpr-GFP) into the growing tube wall enhanced tube length and germination, consistent with a role of TGase as a modulator of cell wall building and strengthening. The secreted pollen TGase catalysed the cross-linking of both PAs (polyamines) into proteins (released by the pollen tube) and His6-Xpr-GFP into endogenous or exogenously added substrates. A similar distribution of TGase activity was observed in planta on pollen tubes germinating inside the style, consistent with a possible additional role for TGase in the interaction between the pollen tube and the style during fertilization.",
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Di Sandro, A, Del Duca, S, Verderio, E, Hargreaves, AJ, Scarpellini, A, Cai, G, Cresti, M, Faleri, C, Iorio, RA, Hirose, S, Furutani, Y, Coutts, IGC, Griffin, M, Bonner, PLR & Serafini-Fracassini, D 2010, 'An extracellular transglutaminase is required for apple pollen tube growth' Biochemical Journal, vol. 429, no. 2, pp. 261-271. https://doi.org/10.1042/BJ20100291

An extracellular transglutaminase is required for apple pollen tube growth. / Di Sandro, Alessia; Del Duca, Stefano; Verderio, Elisabetta; Hargreaves, Alan J.; Scarpellini, Alessandra; Cai, Giampiero; Cresti, Mauro; Faleri, Claudia; Iorio, Rosa A.; Hirose, Shigehisa; Furutani, Yutaka; Coutts, Ian G.C.; Griffin, Martin; Bonner, Philip L.R.; Serafini-Fracassini, Donatella.

In: Biochemical Journal, Vol. 429, No. 2, 05.05.2010, p. 261-271.

Research output: Contribution to journalArticle

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T1 - An extracellular transglutaminase is required for apple pollen tube growth

AU - Di Sandro, Alessia

AU - Del Duca, Stefano

AU - Verderio, Elisabetta

AU - Hargreaves, Alan J.

AU - Scarpellini, Alessandra

AU - Cai, Giampiero

AU - Cresti, Mauro

AU - Faleri, Claudia

AU - Iorio, Rosa A.

AU - Hirose, Shigehisa

AU - Furutani, Yutaka

AU - Coutts, Ian G.C.

AU - Griffin, Martin

AU - Bonner, Philip L.R.

AU - Serafini-Fracassini, Donatella

PY - 2010/5/5

Y1 - 2010/5/5

N2 - An extracellular form of the calcium-dependent protein-cross-linking enzyme TGase (transglutaminase) was demonstrated to be involved in the apical growth of Malus domestica pollen tube. Apple pollen TGase and its substrates were co-localized within aggregates on the pollen tube surface, as determined by indirect immunofluorescence staining and the in situ cross-linking of fluorescently labelled substrates. TGase-specific inhibitors and an anti-TGase monoclonal antibody blocked pollen tube growth, whereas incorporation of a recombinant fluorescent mammalian TGase substrate (histidine-tagged green fluorescent protein: His6-Xpr-GFP) into the growing tube wall enhanced tube length and germination, consistent with a role of TGase as a modulator of cell wall building and strengthening. The secreted pollen TGase catalysed the cross-linking of both PAs (polyamines) into proteins (released by the pollen tube) and His6-Xpr-GFP into endogenous or exogenously added substrates. A similar distribution of TGase activity was observed in planta on pollen tubes germinating inside the style, consistent with a possible additional role for TGase in the interaction between the pollen tube and the style during fertilization.

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KW - germination

KW - green fluorescent proteins

KW - immunohistochemistry

KW - malus

KW - biological models

KW - genetically modified plants

KW - pollen tube

KW - recombinant proteins

KW - substrate specificity

KW - transglutaminases

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Di Sandro A, Del Duca S, Verderio E, Hargreaves AJ, Scarpellini A, Cai G et al. An extracellular transglutaminase is required for apple pollen tube growth. Biochemical Journal. 2010 May 5;429(2):261-271. https://doi.org/10.1042/BJ20100291