title: Adsorption of GST-PI3Kgamma at the air-buffer interface and at substrate and nonsubstrate phospholipid monolayers.
abstract: The recruitment of phosphoinositide 3-kinase gamma (PI3Kgamma) to the cell membrane is a crucial requirement for the initiation of inflammation cascades by second-messenger production. In addition to identifying other regulation pathways, it has been found that PI3Kgamma is able to bind phospholipids directly. In this study, the adsorption behavior of glutathione S-transferase (GST)-PI3Kgamma to nonsubstrate model phospholipids, as well as to commercially available substrate inositol phospholipids (phosphoinositides), was investigated by use of infrared reflection-absorption spectroscopy (IRRAS). The nonsubstrate phospholipid monolayers also yielded important information about structural requirements for protein adsorption. The enzyme did not interact with condensed zwitterionic or anionic monolayers; however, it could penetrate into uncompressed fluid monolayers. Compression to values above its equilibrium pressure led to a squeezing out and desorption of the protein. Protein affinity for the monolayer surface increased considerably when the lipid had an anionic headgroup and contained an arachidonoyl fatty acyl chain in sn-2 position. Similar results on a much higher level were observed with substrate phosphoinositides. No structural response of GST-PI3Kgamma to lipid interaction was detected by IRRAS. On the other hand, protein adsorption caused a condensing effect in phosphoinositide monolayers. In addition, the protein reduced the charge density at the interface probably by shifting the pK values of the phosphate groups attached to the inositol headgroups. Because of their strongly polar headgroups, an interaction of the inositides with the water molecules of the subphase can be expected. This interaction is disturbed by protein adsorption, causing the ionization state of the phosphates to change.
authors.affiliation: Interface Department, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Hermelink
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kirsch
authors.forename: Reinhard
authors.initials: R
authors.lastname: Klinger
authors.forename: Gerald
authors.initials: G
authors.lastname: Reiter
authors.forename: Gerald
authors.initials: G
authors.lastname: Brezesinski
mesh_terms: D000042:Absorption; D000327:Adsorption; D000388:Air; D000818:Animals; D016718:Arachidonic Acid / Q000737:chemistry / Q000378:metabolism; D002021:Buffers; D002460:Cell Line; D058544:Class Ib Phosphatidylinositol 3-Kinase; D005982:Glutathione Transferase / Q000737:chemistry* / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D009200:Myocardial Contraction; D019869:Phosphatidylinositol 3-Kinases / Q000737:chemistry* / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D015398:Signal Transduction; D013055:Spectrophotometry, Infrared
chemical_list: D002021:Buffers; D007527:Isoenzymes; D018129:Phosphatidylinositol Phosphates; D010743:Phospholipids; C060974:phosphatidylinositol 3,4,5-triphosphate; D016718:Arachidonic Acid; D005982:Glutathione Transferase; D019869:Phosphatidylinositol 3-Kinases; D058544:Class Ib Phosphatidylinositol 3-Kinase