A conserved sequence immediately N-terminal to the Bateman domains in AMP-activated protein kinase γ subunits is required for the interaction with the β subunits

TY - JOUR

T1 - A conserved sequence immediately N-terminal to the Bateman domains in AMP-activated protein kinase γ subunits is required for the interaction with the β subunits

AU - Viana, Rosa

AU - Towler, Mhairi C.

AU - Pan, David A.

AU - Carling, David

AU - Viollet, Benoit

AU - Hardie, D. Grahame

AU - Sanz, Pascual

PY - 2007/6

Y1 - 2007/6

N2 - Mammalian AMP-activated protein kinase is a serine/threonine protein kinase that acts as a sensor of cellular energy status. AMP-activated protein kinase is a heterotrimer of three different subunits, i.e. α, β, and γ, with α being the catalytic subunit and β and γ having regulatory roles. Although several studies have defined different domains in α and β involved in the interaction with the other subunits of the complex, little is known about the regions of the γ subunits involved in these interactions. To study this, we have made sequential deletions from the Ntermini of the γ subunit isoforms and studied the interactions with α and β subunits, both by two-hybrid analysis and by coimmunoprecipitation. Our results suggest that a conserved region of 20-25 amino acids in γ1, γ2, and γ3, immediately N-terminal to the Bateman domains, is required for the formation of a functional, active αβγ complex. This region is required for the interaction with the β subunits. The interaction between the α and γ subunits does not require this region and occurs instead within the Bateman domains of the γ subunit, although the α-γ interaction does appear to stabilize the β-γ interaction. In addition, sequential deletions from the C termini of the γ subunits indicate that deletion of any of the CBS (cystathionine β-synthase) motifs prevents the formation of a functional complex with the α and β subunits.

AB - Mammalian AMP-activated protein kinase is a serine/threonine protein kinase that acts as a sensor of cellular energy status. AMP-activated protein kinase is a heterotrimer of three different subunits, i.e. α, β, and γ, with α being the catalytic subunit and β and γ having regulatory roles. Although several studies have defined different domains in α and β involved in the interaction with the other subunits of the complex, little is known about the regions of the γ subunits involved in these interactions. To study this, we have made sequential deletions from the Ntermini of the γ subunit isoforms and studied the interactions with α and β subunits, both by two-hybrid analysis and by coimmunoprecipitation. Our results suggest that a conserved region of 20-25 amino acids in γ1, γ2, and γ3, immediately N-terminal to the Bateman domains, is required for the formation of a functional, active αβγ complex. This region is required for the interaction with the β subunits. The interaction between the α and γ subunits does not require this region and occurs instead within the Bateman domains of the γ subunit, although the α-γ interaction does appear to stabilize the β-γ interaction. In addition, sequential deletions from the C termini of the γ subunits indicate that deletion of any of the CBS (cystathionine β-synthase) motifs prevents the formation of a functional complex with the α and β subunits.

UR - https://www.scopus.com/pages/publications/34447512615

U2 - 10.1074/jbc.M611804200

DO - 10.1074/jbc.M611804200

M3 - Article

C2 - 17403675

AN - SCOPUS:34447512615

SN - 0021-9258

VL - 282

SP - 16117

EP - 16125

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 22

ER -