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Phosphoinositide 3-kinase γ has Multiple Phospholipid Binding Sites

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Abstract

Phosphoinositide 3-kinase γ is a multifunctional enzyme with lipid and protein kinase activities that also acts as a scaffold protein in many diverse signalling processes. The enzyme contains five different domains, but their individual contributions to membrane binding are not fully understood. Here, using in vitro liposome binding assays of individual domains and deletion constructs of human phosphoinositide 3-kinase γ, we show that each domain is capable of binding anionic phospholipids to varying degrees, depending on the charge of the anionic substrate. Moreover, with the exception of the C2-domain, deletion of any single protein domain results in a complete loss of kinase activity toward both lipids and proteins.

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Abbreviations

PI3 Kγ:

Phosphoinositide 3-kinase γ

NtD:

N-terminal domain (the first N-terminal 219 aminoacids of PI3 Kγ)

RBD:

Ras binding domain

C2D:

C2 domain

helD:

Helical domain

catD:

Catalytic domain

NtD-Del:

Deletion mutant, where NtD is missing

RBD-Del:

Deletion mutant, where RBD is missing

C2D-Del:

Deletion mutant, where C2D is missing

helD-Del:

Deletion mutant, where helD is missing

PtdIns:

Phosphatidylinositol

PtdIns(4)P:

Phosphatidylinositol-4-phosphate

PtdIns(4,5)P2 :

Phosphatidylinositol-4,5-bisphosphate

PtdSer:

Phosphatidylserine

PA:

Phosphatidate

Rh-PtdEtn:

Rhodamine-PtdEtn

Chol:

Cholesterol

Pam:

Palmitic acid

Ste:

Stearic acid

Δ4Ach:

Arachidonic acid

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Authors and Affiliations

  1. Universitätsklinikum Jena, Institut für Biochemie II, Nonnenplan 2, 07740, Jena, Germany

    Carsten Schmidt, Margret Schilli-Westermann, Reinhard Klinger & Cornelia Kirsch

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  1. Carsten Schmidt
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  2. Margret Schilli-Westermann
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  3. Reinhard Klinger
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  4. Cornelia Kirsch
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Correspondence to Carsten Schmidt.

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Schmidt, C., Schilli-Westermann, M., Klinger, R. et al. Phosphoinositide 3-kinase γ has Multiple Phospholipid Binding Sites. Protein J 29, 127–135 (2010). https://doi.org/10.1007/s10930-010-9232-x

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