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Substrate specificity of Pasteurella multocida toxin for α subunits of heterotrimeric G proteins
Author(s) -
Orth Joachim H. C.,
Fester Ines,
Siegert Peter,
Weise Markus,
Lanner Ulrike,
Kamitani Shigeki,
Tachibana Taro,
Wilson Brenda A.,
Schlosser Andreas,
Horiguchi Yasuhiko,
Aktories Klaus
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.12-213900
Subject(s) - heterotrimeric g protein , protein subunit , g protein , toxin , biology , g beta gamma complex , fusion protein , virulence factor , gs alpha subunit , virulence , microbiology and biotechnology , biochemistry , signal transduction , recombinant dna , gene
Pasteurella multocida is the causative agent of a number of epizootic and zoonotic diseases. Its major virulence factor associated with atrophic rhinitis in animals and dermonecrosis in bite wounds is P. multocida toxin (PMT). PMT stimulates signal transduction pathways downstream of heterotrimeric G proteins, leading to effects such as mitogenicity, blockade of apoptosis, or inhibition of osteoblast differentiation. On the basis of Gα i2 , it was demonstrated that the toxin deamidates an essential glutamine residue of the Gα i2 subunit, leading to constitutive activation of the G protein. Here, we studied the specificity of PMT for its G‐protein targets by mass spectrometric analyses and by utilizing a monoclonal antibody, which recognizes specifically G proteins deamidated by PMT. The studies revealed deamidation of 3 of 4 families of heterotrimeric G proteins (Gα q/11 , Gα i1,2,3 , and Gα 12/13 of mouse or human origin) by PMT but not by a catalytic inactive toxin mutant. With the use of G‐protein fragments and chimeras of responsive or unresponsive G proteins, the structural basis for the discrimination of heterotrimeric G proteins was studied. Our results elucidate substrate specificity of PMT on the molecular level and provide evidence for the underlying structural reasons of substrate discrimination.—Orth, J. H. C., Fester, I., Siegert, P., Weise, M., Lanner, U., Kamitani, S., Tachibana, T, Wilson, B. A., Schlosser, A., Horiguchi, Y., Aktories, K. Substrate specificity of Pasteurella multocida toxin for α subunits of heterotrimeric G proteins. FASEB J. 27, 832–842 (2013). www.fasebj.org