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Molecular cloning, expression and characterization of three distinctive genes encoding methionine aminopeptidases in cyanobacterium Synechocystis sp. strain PCC6803

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Abstract

Methionine aminopeptidase, known to be encoded by single genes in prokaryotes, is a cobalt-dependent enzyme that catalyzes the removal of N-terminal methionine residues from nascent polypeptides. Three ORFs encoding putative methionine aminopeptidases from the genome of cyanobacterium Synechocystis sp. strain PCC6803, designated as slr0786 (map-1), slr0918 (map-2) and sll0555 (map-3) were cloned and expressed in Escherichia coli. The purified recombinant proteins encoded by map-1 and map-3 had much higher methionine aminopeptidase activity than the recombinant protein encoded by map-2. Comparative analysis revealed that the three recombinant enzymes differed in their substrate specificity, divalent ion requirement, pH, and temperature optima. The broad activities of the iso-enzymes are discussed in light of the structural similarities with other peptidase families and their levels of specificity in the cell. Potential application of cyanobacterial MetAPs in the production of recombinant proteins used in medicine is proposed. This is the first report of a prokaryote harboring multiple methionine aminopeptidases.

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Abbreviations

map :

Gene encoding methionine aminopeptidase

MetAP :

Methionine aminopeptidase

eMetAP-Ia :

Escherichia coli methionine aminopeptidase type Ia

yMetAP-Ib :

Yeast methionine aminopeptidase type Ib

yMetAP-IIa :

Yeast methionine aminopeptidase type IIa

hMetAP-IIb :

Human methionine aminopeptidase type IIb

pfMetAP−IIa :

Pyrococcus furiosis methionine aminopeptidase type Ia

bst MetAP-Ia :

Bacillus stearothermophilus methionine aminopeptidase type Ia

c1MetAP-Ia :

Cyanobacterial methionine aminopeptidase type Ia encoded by map-1

c2MetAP-Ia :

Cyanobacterial methionine aminopeptidase type Ia encoded by map-2

c3MetAP-Ib :

Cyanobacterial methionine aminopeptidase type Ib, ncoded by map-3

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Acknowledgements

This work was supported by a fellowship from the Japanese Society for the Promotion of Science and partly by a grant from the Bulgarian National Research Fund (K-609/1997). We thank Professor Deborah Zamble, Alistair Dias, Daria Yu, Teodora Jivkova, Asuka Mase, Nickolay Tzvetkov, Svetoslav Dimov, Dr. Igor Splawski and Dr. Michinori Mutsuda for help and discussions.

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Authors and Affiliations

  1. Institute of Molecular Biology, Bulgarian Academy of Sciences, bl. 21 Acad. G. Bonchev St., 1113 , Sofia, Bulgaria

    Anelia Atanassova, Tamara Pajpanova & Ivan Ivanov

  2. Laboratory of Plant Molecular Biology and Biological Informatics, Nagoya University, Graduate School of Human Informatics, 464 – 8601, Nagoya, Japan

    Mamoru Sugita

  3. Graduate School of Natural Sciences, Nagoya City University, 467 – 8501, Nagoya , Japan

    Masahiro Sugiura

  4. Department of Chemistry, University of Toronto, 80 St. George St., Lash Miller Chemical Laboratories, Toronto, Ontario , M5S 3H6, Canada

    Anelia Atanassova

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  1. Anelia Atanassova
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Correspondence to Anelia Atanassova.

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Atanassova, A., Sugita, M., Sugiura, M. et al. Molecular cloning, expression and characterization of three distinctive genes encoding methionine aminopeptidases in cyanobacterium Synechocystis sp. strain PCC6803. Arch Microbiol 180, 185–193 (2003). https://doi.org/10.1007/s00203-003-0576-x

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  • DOI: https://doi.org/10.1007/s00203-003-0576-x

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