Functional expression and characterisation of a new human phosphatidylinositol 4-kinase PI4K230.
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Functional expression and characterisation of a new human phosphatidylinositol 4-kinase PI4K230. / Gehrmann, T; Gülkan, H; Suer, S; Herberg, F W; Balla, A; Vereb, G; Mayr, Georg W.; Heilmeyer, L M.
In: BBA-BIOMEMBRANES, Vol. 1437, No. 3, 3, 1999, p. 341-356.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Functional expression and characterisation of a new human phosphatidylinositol 4-kinase PI4K230.
AU - Gehrmann, T
AU - Gülkan, H
AU - Suer, S
AU - Herberg, F W
AU - Balla, A
AU - Vereb, G
AU - Mayr, Georg W.
AU - Heilmeyer, L M
PY - 1999
Y1 - 1999
N2 - By constructing DNA probes we have identified and cloned a human PtdIns 4-kinase, PI4K230, corresponding to a mRNA of 7.0 kb. The cDNA encodes a protein of 2044 amino acids. The C-terminal part of ca. 260 amino acids represents the catalytic domain which is highly conserved in all recently cloned PtdIns 4-kinases. N-terminal motifs indicate multiple heterologous protein interactions. Human PtdIns 4-kinase PI4K230 expressed in vitro exhibits a specific activity of 58 micromol mg-1min-1. The enzyme expressed in Sf9 cells is essentially not inhibited by adenosine, it shows a high Km for ATP of about 300 microM and it is half-maximally inactivated by approximately 200 nM wortmannin. These data classify this enzyme as type 3 PtdIns 4-kinase. Antibodies raised against the N-terminal part moderately activate and those raised against the C-terminal catalytic domain inhibit the enzymatic activity. The coexistence of two different type 3 PtdIns 4-kinases, PI4K92 and PI4K230, in several human tissues, including brain, suggests that these enzymes are involved in distinct basic cellular functions.
AB - By constructing DNA probes we have identified and cloned a human PtdIns 4-kinase, PI4K230, corresponding to a mRNA of 7.0 kb. The cDNA encodes a protein of 2044 amino acids. The C-terminal part of ca. 260 amino acids represents the catalytic domain which is highly conserved in all recently cloned PtdIns 4-kinases. N-terminal motifs indicate multiple heterologous protein interactions. Human PtdIns 4-kinase PI4K230 expressed in vitro exhibits a specific activity of 58 micromol mg-1min-1. The enzyme expressed in Sf9 cells is essentially not inhibited by adenosine, it shows a high Km for ATP of about 300 microM and it is half-maximally inactivated by approximately 200 nM wortmannin. These data classify this enzyme as type 3 PtdIns 4-kinase. Antibodies raised against the N-terminal part moderately activate and those raised against the C-terminal catalytic domain inhibit the enzymatic activity. The coexistence of two different type 3 PtdIns 4-kinases, PI4K92 and PI4K230, in several human tissues, including brain, suggests that these enzymes are involved in distinct basic cellular functions.
M3 - SCORING: Zeitschriftenaufsatz
VL - 1437
SP - 341
EP - 356
JO - BBA-BIOMEMBRANES
JF - BBA-BIOMEMBRANES
SN - 0005-2736
IS - 3
M1 - 3
ER -
