Biochemical characterization of two analogues of the apoptosis-linked gene 2 protein in Dictyostelium discoideum and interaction with a physiological partner in mammals, murine Alix

The Journal of Biological Chemistry
Laurence AubryGérard Klein

Abstract

Two homologues, Dd-ALG-2a and Dd-ALG-2b, of the mammalian calcium-binding protein ALG-2 (apoptosis-linked gene 2) have been characterized in the cellular slime mold Dictyostelium discoideum. Fluorescence titrations showed that both proteins bind calcium ions with affinities (Ca2+)(0.5) of 30 and 450 microm, respectively, at sites specific to calcium. Calcium ion binding resulted in changes of conformation associated with the unmasking of hydrophobic regions of the proteins. Surface plasmon resonance analysis showed that Dd-ALG-2a homodimers formed (K(D) of 1 microm) at calcium ion concentrations similar to those necessary for Ca2+-induced conformational changes. Deletion of the hydrophobic N-terminal sequence or EF-hand 5 of Dd-ALG-2a prevented dimerization. The Dd-ALG-2b homodimer was not detected, and the Dd-ALG-2a/2b heterodimer formed only when Dd-ALG-2b was the immobilized partner. Murine Alix formed a heterodimer (K(D) = 0.6 microm) with Dd-ALG-2a but not with Dd-ALG-2b, and the interaction strictly depended upon calcium ions. The DeltaNter construct of Dd-ALG-2a lost its interaction capacity with mouse Alix. The genes encoding both proteins, Dd-alg-2a and -2b, were expressed in growing cells. The levels of mRNA were at a...Continue Reading

References

Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J L Dynes, R A Firtel
Jan 1, 1987·Methods in Cell Biology·W NellenR A Firtel
Jan 1, 1994·Methods in Cell Biology·D M BersR Nuccitelli
Jan 1, 1996·Methods in Enzymology·D G HigginsT J Gibson
Feb 20, 1999·The Journal of Biological Chemistry·S CheJ Kuang
Jan 3, 2001·Proceedings of the National Academy of Sciences of the United States of America·A H Wyllie, P Golstein
Jun 9, 2001·Methods in Cell Biology·J P LevraudP Golstein
Jan 17, 2002·Functional & Integrative Genomics·A KuspaG Shaulsky
May 1, 1960·Proceedings of the National Academy of Sciences of the United States of America·W F Whittingham, K B Raper

❮ Previous
Next ❯

Citations

Nov 26, 2002·Biochimica Et Biophysica Acta·Masatoshi MakiHideki Shibata
Nov 26, 2002·Biochimica Et Biophysica Acta·Joachim KrebsRosmarie Caduff
Dec 24, 2010·PloS One·Dorian GuettaLaurence Aubry
Feb 2, 2010·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Irina V Shemarova
Sep 1, 2004·Biochemical and Biophysical Research Communications·Lalita Subramanian, Arthur S Polans
Jun 13, 2006·European Journal of Cell Biology·Sara MatteiLaurence Aubry
Jun 10, 2006·European Journal of Cell Biology·Marija MarkoAngelika A Noegel
Feb 15, 2005·Developmental Biology·Sara MatteiLaurence Aubry
Jan 21, 2004·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Feb 15, 2020·International Journal of Molecular Sciences·Danton H O'DayMichael A Myre
Aug 18, 2012·The Journal of Experimental Biology·Camille Martinand-MariStephen Baghdiguian

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.