Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme

F Meggio, B Boldyreff, O Marin, L A Pinna, O G Issinger

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Udgivelsesdato: 1992-Feb-15
OriginalsprogEngelsk
TidsskriftEuropean Journal of Biochemistry
Vol/bind204
Udgave nummer1
Sider (fra-til)293-297
Antal sider4
ISSN0014-2956
StatusUdgivet - 15. feb. 1992

Fingeraftryk

Casein Kinase II
Holoenzymes
Peptides
Catalytic Domain
Acidic Amino Acids
Urea
Enzymes

Citer dette

@article{b14c86505a4111de839d000ea68e967b,
title = "Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme",
abstract = "Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce the loss of kinase activity, strongly suggesting a protective function for the beta subunit. Assaying different peptides for specificity toward the recombinant alpha subunit and the recombinant reconstituted enzyme, showed that the presence of the beta subunit could modify the specificity of the catalytic alpha subunit. Therefore, a dual function for the beta subunit is proposed which confers both specificity and stability to the catalytic alpha subunit within the CK-2 holoenzyme complex. The peptide DLEPDEELEDNPNQSDL, reproducing the highly acidic amino acid 55-71 segment of the human beta subunit, counteracts the stimulatory effect of the beta subunit on the alpha subunit activity and partially substitutes the beta subunit in conferring thermal stability to the alpha subunit. No such effect is induced by the peptide MSSSEEVSW, reproducing the N-terminal segment of the beta subunit including the autophosphorylation site. It is suggested that the acidic domain of the beta subunit, encompassing residues 55-71, plays a role in the interactions between the beta and alpha subunits.",
keywords = "Amino Acid Sequence, Animals, Casein Kinases, Enzyme Stability, Hot Temperature, Kinetics, Liver, Macromolecular Substances, Magnesium, Molecular Sequence Data, Protein Kinases, Rats, Recombinant Proteins, Substrate Specificity, Urea",
author = "F Meggio and B Boldyreff and O Marin and Pinna, {L A} and Issinger, {O G}",
year = "1992",
month = "2",
day = "15",
language = "English",
volume = "204",
pages = "293--297",
journal = "European Journal of Biochemistry",
issn = "0014-2956",
publisher = "AAAI Press",
number = "1",

}

Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme. / Meggio, F; Boldyreff, B; Marin, O; Pinna, L A; Issinger, O G.

I: European Journal of Biochemistry, Bind 204, Nr. 1, 15.02.1992, s. 293-297.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme

AU - Meggio, F

AU - Boldyreff, B

AU - Marin, O

AU - Pinna, L A

AU - Issinger, O G

PY - 1992/2/15

Y1 - 1992/2/15

N2 - Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce the loss of kinase activity, strongly suggesting a protective function for the beta subunit. Assaying different peptides for specificity toward the recombinant alpha subunit and the recombinant reconstituted enzyme, showed that the presence of the beta subunit could modify the specificity of the catalytic alpha subunit. Therefore, a dual function for the beta subunit is proposed which confers both specificity and stability to the catalytic alpha subunit within the CK-2 holoenzyme complex. The peptide DLEPDEELEDNPNQSDL, reproducing the highly acidic amino acid 55-71 segment of the human beta subunit, counteracts the stimulatory effect of the beta subunit on the alpha subunit activity and partially substitutes the beta subunit in conferring thermal stability to the alpha subunit. No such effect is induced by the peptide MSSSEEVSW, reproducing the N-terminal segment of the beta subunit including the autophosphorylation site. It is suggested that the acidic domain of the beta subunit, encompassing residues 55-71, plays a role in the interactions between the beta and alpha subunits.

AB - Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce the loss of kinase activity, strongly suggesting a protective function for the beta subunit. Assaying different peptides for specificity toward the recombinant alpha subunit and the recombinant reconstituted enzyme, showed that the presence of the beta subunit could modify the specificity of the catalytic alpha subunit. Therefore, a dual function for the beta subunit is proposed which confers both specificity and stability to the catalytic alpha subunit within the CK-2 holoenzyme complex. The peptide DLEPDEELEDNPNQSDL, reproducing the highly acidic amino acid 55-71 segment of the human beta subunit, counteracts the stimulatory effect of the beta subunit on the alpha subunit activity and partially substitutes the beta subunit in conferring thermal stability to the alpha subunit. No such effect is induced by the peptide MSSSEEVSW, reproducing the N-terminal segment of the beta subunit including the autophosphorylation site. It is suggested that the acidic domain of the beta subunit, encompassing residues 55-71, plays a role in the interactions between the beta and alpha subunits.

KW - Amino Acid Sequence

KW - Animals

KW - Casein Kinases

KW - Enzyme Stability

KW - Hot Temperature

KW - Kinetics

KW - Liver

KW - Macromolecular Substances

KW - Magnesium

KW - Molecular Sequence Data

KW - Protein Kinases

KW - Rats

KW - Recombinant Proteins

KW - Substrate Specificity

KW - Urea

M3 - Journal article

C2 - 1740140

VL - 204

SP - 293

EP - 297

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 1

ER -