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Functional analysis of CK2β-derived syntheticfragments

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Abstract

Synthetic peptides reproducing the amino and carboxyl terminal region of CK2 β subunit have been analyzed for their ability to mimic different properties of full length β subunit. Peptide β[1-77], containing both the autophosphorylation site and the down-regulatory domain 55-64, is readily phosphorylated by a subunit whose activity is concomitantly inhibited. Such inhibition is accompanied by a weak interaction detectable by BIAcore sensograms but not by far Western blots, and is not reversed by polylysine which conversely overcome inhibition of calmodulin phosphorylation by full length β subunit. A strong interaction with α is observed with β[155-215] but not with its shorter derivative β[170-215] as judged from far Western blotting and sucrose gradient ultracentrifugation analysis. Both peptides, however, affect the regular interaction between α and β subunits altering the autophosphorylation pattern and responsiveness to salt. β[155-215], unlike β[170-215] tends to aggregate more readily than full length β subunit. This behaviour which is reminiscent of the homodimerization of full length β subunit, would indicate that tight self-association of β[155-215] crucially depends on residues in the 155-170 sequence. Failure of β[1-77] fragment to mediate responsiveness to polybasic peptides and accentuated self-association propensity of β[155-215] suggest that other structural elements between the sequences 1-77 and 155-215 are required in order to confer optimal functionality to the β subunit.

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Authors and Affiliations

  1. Dipartimento di Chimica Biologica, CNR, Centro di Studio delle Biomembrane, Università di Padova, Italy

    Flavio Meggio, Stefania Sarno & Lorenzo A. Pinna

  2. CRIBI Biotechnology Center, Padova, Italy

    Oriano Marin

Authors
  1. Flavio Meggio
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  2. Oriano Marin
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  3. Stefania Sarno
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  4. Lorenzo A. Pinna
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Meggio, F., Marin, O., Sarno, S. et al. Functional analysis of CK2β-derived syntheticfragments. Mol Cell Biochem 191, 35–42 (1999). https://doi.org/10.1023/A:1006822632216

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