Department of Biological and Environmental Sciences

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Browsing Department of Biological and Environmental Sciences by Author "Lin Liu"

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    Characterization of -soluble n-ethylmaleimide–sensitive fusion attachment protein in alveolar type ii cells implications in lung surfactant secretion
    (2002) Abonyo, B. O.; Wang, Pengcheng; Narasaraju, Telugu A.; Rowan III, William H.; Zimmerman, Un-Jin; Lin Liu
    N-ethylmaleimide–sensitive fusion protein (NSF) and soluble NSF attachment protein (α-SNAP) are thought to be soluble factors that transiently bind and disassemble SNAP receptor complex during exocytosis in neuronal and endocrine cells. Lung surfactant is secreted via exocytosis of lamellar bodies from alveolar epithelial type II cells. However, the secretion of lung surfactant is a relatively slow process, and involvement of SNAP receptor and its cofactors (NSF and α-SNAP) in this process has not been demonstrated. In this study, we investigated a possible role of α-SNAP in surfactant secretion. α-SNAP was predominantly associated with the membranes in alveolar type II cells as determined by Western blot and immunocytochemical analysis using confocal microscope. Membrane-associated α-SNAP was not released from the membrane fraction when the cells were lyzed in the presence of Ca2+ or Mg2+ATP. The alkaline condition (0.1 M Na2CO3, pH 12), known to extract peripheral membrane proteins also failed to release it from the membrane. Phase separation using Triton X-114 showed that α-SNAP partitioned into both aqueous and detergent phases. NSF had membrane-bound characteristics similar to α-SNAP in type II cells. Permeabilization of type II cells with β-escin resulted in a partial loss of α-SNAP from the cells, but cellular NSF was relatively unchanged. Addition of exogenous α-SNAP to the permeabilized cells increased surfactant secretion in a dose-dependent manner, whereas exogenous NSF has much less effects. An α-SNAP antisense oligonucleotide decreased its protein level and inhibited surfactant secretion. Our results suggest a role of α-SNAP in lung surfactant secretion.
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    Syntaxin 2 and SNAP-23 are required for regulated surfactant secretion
    (2014) Abonyo, B. O.; Wang, Pengcheng; Lin Liu
    The secretion of lung surfactant in alveolar type II cells is a complex process involving the fusion of lamellar bodies with the plasma membrane. This process is somewhat different from the exocytosis of hormones and neurotransmitters. For example, it is a relatively slower process, and lamellar bodies are very large vesicles with a diameter of approximately 1 microm. SNARE proteins are the conserved molecular machinery of exocytosis in the majority of secretory cells. However, their involvement in surfactant secretion has not been reported. Here, we showed that syntaxin 2 and SNAP-23 are expressed in alveolar type II cells. Both proteins are associated with the plasma membrane, and to some degree with lamellar bodies. An antisense oligonucleotide complementary to syntaxin 2 decreased its mRNA and protein levels. The same oligonucleotide also inhibited surfactant secretion, independent of secretagogues. A peptide derived from the N-terminus of syntaxin 2 or the C-terminus of SNAP-23 significantly inhibited Ca(2+)- and GTPgammaS-stimulated surfactant secretion from permeabilized type II cells in a dose-dependent manner. Furthermore, introduction of anti-syntaxin 2 or anti-SNAP-23 antibodies into permeabilized type II cells also inhibited surfactant release. Our results suggest that syntaxin 2 and SNAP-23 are required for regulated surfactant secretion.