Differential expression of a C-terminal splice variant of phosphatidylinositol transfer protein beta lacking the constitutive-phosphorylated Ser262 that localizes to the Golgi compartment

Mammalian PITPbeta (phosphatidylinositol transfer protein beta) is a 272-amino-acid polypeptide capable of transferring PtdIns, PtdCho and SM (sphingomyelin) between membrane bilayers. It has been reported that Ser262 present in the C-terminus of PITPbeta is constitutively phosphorylated and determines Golgi localization. We provide evidence for the expression of an sp (splice) variant of PITPbeta (PITPbeta-sp2) where the C-terminal 15 amino acids of PITPbeta-sp1 are replaced by an alternative C-terminus of 16 amino acids. PITPbeta-sp1 is the product of the first 11 exons, whereas PITPbeta-sp2 is a product of the first 10 exons followed by the twelfth exon--exon 11 being 'skipped'. Both splice variants are capable of PtdIns and PtdCho transfer, with PITPbeta-sp2 being unable to transport SM. PITPbeta is ubiquitously expressed, with the highest amounts of PITPbeta found in HL60 cells and in rat liver; HL60 cells express only PITPbeta-sp1, whereas rat liver expresses both sp variants in similar amounts. In both cell types, PITPbeta-sp1 is constitutively phosphorylated and both the PtdIns and PtdCho forms of PITPbeta-sp1 are present. In contrast, PITPbeta-sp2 lacks the constitutively phosphorylated Ser262 (replaced with glutamine). Nonetheless, both PITPbeta variants localize to the Golgi and, moreover, dephosphorylation of Ser262 of PITPbeta-sp1 does not affect its Golgi localization. The presence of PITPbeta sp variants adds an extra level of proteome complexity and, in rat liver, the single gene for PITPbeta gives rise to seven distinct protein species that can be resolved on the basis of their charge differences.     

Phosphatidylinositol transfer proteins: a requirement in signal transduction and vesicle traffic

Phosphatidylinositol transfer protein (PITP) was originally identified and named because of its ability to transport phosphatidylinositol through the aqueous phase from one membrane compartment to another. Recent data, however, indicate unanticipated roles for PITP in the coupling of PIP₂ synthesis to signal transduction reactions and to membrane traffic in mammalian cells. PITP was recently purified on the basis of its ability to restore cellular functions in permeabilized cells depleted of cytosolic proteins. These functions include cell-surface receptor-regulated hydrolysis of PIP₂ by phospholipases Cβ- and γ-isozymes, regulated release of secretory granules, and the budding of constitutive secretory vesicles and immature secretory granules from the trans-Golgi network. In the yeast Saccharomyces cerevisiae, a PITP was identified from a mutant strain with a defect in the secretory pathway (SEC14) and therefore required for cell viability; in Yarrowia lipolytica, PITP is required for differentiation from a yeast to a mycelial growth form. We are just beginning to unravel the intriguing mechanisms by which PITP/SEC14 may accomplish its function in eukaryotic cells in signal transduction and membrane trafficking. BioEssays 20 :423-432, 1998. © 1998 John Wiley & Sons Inc.     

Phosphatidic acid regulation of phosphatidylinositol 4-phosphate 5-kinases

Phosphatidic acid (PA) production by receptor-stimulated phospholipase D is believed to play an important role in the regulation of cell function. The second messenger function of PA remains to be elucidated. PA can bind and affect the activities of different enzymes and here we summarise the current status of activation of Type I phosphatidylinositol 4-phosphate 5-kinase by PA. Type 1 phosphatidylinositol 4-phosphate 5-kinase is also regulated by ARF proteins as is phospholipase D and we discuss the contributions of ARF and PA towards phosphatidylinositol(4,5)bisphosphate synthesis at the plasma membrane.     

Coupling Efficiency of Surface Plasmon Polaritons for 1D Plasmonic Gratings: Role of Under- and Over-Milling

This paper reports the successful excitation of surface plasmon polaritons (SPPs) through 1D metallic grating on higher refractive index GaP substrate. Coupling efficiency (η) of a free-space transverse-magnetic (TM) plane-wave mode into a SPP mode is crucial for many plasmonic devices. This η predominantly depends on the fabrication (milling) parameters and the factors (under- and over-milling) affecting the η is investigated experimentally and numerically. First of all, η is estimated by measuring the transmission spectra obtained through the plasmonic grating structures by varying the slit width (a) for a fixed period (Λ) and the thickness (t) of the gold (Au) film in which the grating is formed. The wave vector of the incident light is tuned to match the wave vector of the SPP, to get maximum η. For an optimum Au film thickness, a slit width of half of the periodicity of 770 nm in the grating device yields a maximum η. Such grating devices support only a fundamental plasmonic mode because the profile/shape of the slit in the grating device is more like a sinusoidal nature. Furthermore, such grating offers intermediate scattering to the incident light and the SPP as well which in-truns couple more incident energy to the SPPs. Moreover, over-milling results in decreased η where the crystalline plane of the substrate is disturbed. Finite element method (FEM) in COMSOL modeling is used to understand the underlying physics. This study is very useful for the development of the device application in real word.     

RETRACTED ARTICLE: Homo and heterologous expression of the HpPKS2 gene in Hypericum perforatum and Bacopa monnieri

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