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A methionine synthase homolog is associated with secretory vesicles in tobacco pollen tubes

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Abstract

Seven isoforms of 85 kDa polypeptides (p85) were identified as methionine synthase (MetE) homologs by partial aminoacid sequencing in tobacco pollen tube extracts. Immunocytochemistry data showed a localization of the antigen on the surface of tip-focussed post-Golgi secretory vesicles (SVs), that appear to be partially associated with microtubules (Mts). The chemical dissection of pollen tube high speed supernatant (HSS) showed that two distinct pools of MetE are present in pollen tubes, one being the more acidic isoforms sedimenting at 15S and the remaining at 4S after zonal centrifugation through a sucrose density gradient. The identification of the MetE within the pollen tube and its possible participation as methyl donor in a wide range of metabolic reactions, makes it a good subject for studies on pollen tube growth regulation.

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Abbreviations

MetE:

Methionine synthase

SVs:

Secretory vesicles

HSS:

High speed supernatant

SAM:

S-adenosylmethionine

PKC:

Protein Kinase C

CK2:

Casein Kinase II

Mts:

Microtubules

VN:

Vegetative nucleus

GC:

Generative cell

KRPs:

Kinesin-related polypeptides

IC:

Intermediate Chains

LC:

Light Chains

MetT:

Pollen tube MetE homolog

Afs:

Actin Filaments

Mab:

Monoclonal Antibody

BSA:

Bovine Serum Albumin

TAME:

Tosyl-L-Arginine Methyl Ester

PMSF:

Phenylmethanesulfonyl Fluoride

SDS-PAGE:

Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis

Tris:

Tris(hydroxymethyl-d3)amino-d2-methane

Pipes:

Piperazine-1,4-bis(2-ethane-sulfonic acid)

DTT:

1,4-Dithio-DL-threitol

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Acknowledgements

We gratefully thanks Prof. Gianni Piperno (Mount Sinai School of Medicine, NY, USA) for providing the anti-p28 antiserum and for the critical reading of the manuscript. Many thanks to Prof. Schroeder (Institute of Biology II, University of Freiburg Germany) and Prof. Hesse (Max Planck Institute of Molecular Plant Physiology, Golm, Germany) for providing the anti-MetE antibodies. We would also express our sincere gratitude to Prof. V.K. Sawhney (University of Saskatchewan, Canada) for his kindness and helpful revision of the manuscript. This work was partially supported by ERBIC 18CT 960072 of the European Commission and by Siena and Milano Universities.

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Authors and Affiliations

  1. Dipartimento di Biologia, Università degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy

    Alessandra Moscatelli

  2. Dipartimento di Scienze Ambientali “G. Sarfatti”, Università degli Studi di Siena, Via P. A. Mattioli 4, 53100, Siena, Italy

    Monica Scali, Rita Vignani, Fabrizio Ciampolini & Mauro Cresti

  3. Department of Biochemistry, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056, Basel, Switzerland

    Cristina Prescianotto-Baschong

  4. Department de Chimie des Proteines, CEA/Grenoble, 17 Rue des Martyrs, 38054, Grenoble Cèdex, France

    Myriam Ferro & Jerome Garin

Authors
  1. Alessandra Moscatelli
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  2. Monica Scali
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  3. Cristina Prescianotto-Baschong
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  4. Myriam Ferro
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  5. Jerome Garin
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  6. Rita Vignani
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  7. Fabrizio Ciampolini
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  8. Mauro Cresti
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Correspondence to Mauro Cresti.

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Moscatelli, A., Scali, M., Prescianotto-Baschong, C. et al. A methionine synthase homolog is associated with secretory vesicles in tobacco pollen tubes. Planta 221, 776–789 (2005). https://doi.org/10.1007/s00425-005-1487-7

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  • DOI: https://doi.org/10.1007/s00425-005-1487-7

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