Isolation and mass spectrometric characterization of an oxidized form of vasostatin I, anN-terminal chromogranin A-derived protein, from bovine chromaffin cells

1995 ◽  
Vol 30 (11) ◽  
pp. 1599-1604 ◽  
Author(s):  
L. Dillen ◽  
X. Y. Zhang ◽  
M. Claeys ◽  
F. Liang ◽  
W. P. De Potter ◽  
...  
Keyword(s):  
Chromogranin A ◽  
Chromaffin Cells ◽  
Mass Spectrometric ◽  
Bovine Chromaffin Cells

scholarly journals Effect of secretagogues on chromogranin A synthesis in bovine cultured chromaffin cells. Possible regulation by protein kinase C

1989 ◽  
Vol 260 (3) ◽  
pp. 915-922 ◽  
Author(s):  
J P Simon ◽  
M F Bader ◽  
D Aunis
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Chromogranin A ◽  
Chromaffin Cells ◽  
Cell Types ◽  
Term Treatment ◽  
Synthesis Rate ◽  
Kinase C ◽  
Long Term Treatment ◽  
Bovine Chromaffin Cells

Chromogranin A is a major component of storage granules in many different secretory cell types. After [35S]methionine labelling of proteins from cultured bovine chromaffin cells, chromogranin A was immunoprecipitated with specific antibodies, and the radioactivity incorporated into chromogranin A was determined and used as an index of its synthesis rate. Depolarization of cells with nicotine or high K+ evoked a Ca2+-dependent increase in chromogranin A synthesis, whereas muscarine, which does not evoke significant Ca2+ influx from bovine chromaffin cells, had no effect on chromogranin A synthesis. Forskolin, an activator of adenylate cyclase, affected neither the basal nor the nicotine-stimulated rate of chromogranin A synthesis. In contrast, 12-O-tetradecanoylphorbol 13-acetate (TPA), an activator of protein kinase C, significantly enhanced the incorporation of radioactivity into chromogranin A. Sphingosine, an inhibitor of protein kinase C, abolished both nicotine-stimulated and TPA-induced chromogranin A synthesis. In addition, long-term treatment of chromaffin cells with TPA decreased protein kinase C activity and inhibited the nicotine-stimulated chromogranin A synthesis. These results suggest that protein kinase C may play an important role in the control of chromogranin A synthesis.

scholarly journals Characterization of Rab3A, Rab3B and Rab3C: different biochemical properties and intracellular localization in bovine chromaffin cells

1997 ◽  
Vol 324 (1) ◽  
pp. 85-90 ◽  
Author(s):  
Chong-Gee LIN ◽  
Yu-Chi LIN ◽  
Hwan-Wun LIU ◽  
Lung-Sen KAO
Keyword(s):  
Subcellular Localization ◽  
Binding Affinity ◽  
Chromaffin Cells ◽  
Intracellular Localization ◽  
Biochemical Properties ◽  
Amino Acid Residues ◽  
C Terminus ◽  
Bovine Chromaffin Cells ◽  
Adrenal Chromaffin

In this study we examined the biochemical properties and subcellular localization of Rab3A, Rab3B and Rab3C in bovine adrenal chromaffin cells. The Kd for guanosine 5′-[γ-thio]triphosphate (GTP[S]) of the three Rab3 proteins was 15, 2700 and 204 nM for Rab3A, Rab3B and Rab3C respectively. The intrinsic GTPase activity of the three Rab3 proteins seemed similar and was increased approx. 3-fold by bovine chromaffin cell lysate. Truncation of the C-terminal 31 amino acid residues decreased the binding affinity for GTP[S] of the three Rab3 proteins. When the C-terminus of Rab3C was replaced with that of Rab3A, the binding affinity of Rab3C for GTP[S] was decreased, but the replacement did not affect the affinity of Rab3B for GTP[S]. Immunostaining experiments showed that Rab3A, Rab3B and Rab3C are localized separately within chromaffin cells. Anti-Rab3A and anti-Rab3C antibodies stained vesicle-like structures, whereas anti-Rab3B antibody distinctly stained the plasma membrane. In summary, bovine chromaffin cells express the three Rab3 proteins but the subcellular localization and biochemical properties of the three Rab3 proteins are distinct.

Pb2(+)-induced secretion from bovine chromaffin cells: fura-2 as a probe for Pb2+

1990 ◽  
Vol 259 (5) ◽  
pp. C762-C768 ◽  
Author(s):  
J. L. Tomsig ◽  
J. B. Suszkiw
Keyword(s):  
Dissociation Constant ◽  
Chromaffin Cells ◽  
Ionic Composition ◽  
Catecholamine Release ◽  
Apparent Dissociation Constant ◽  
Fura 2 ◽  
Bovine Chromaffin Cells ◽  
In Cells

The effect of Pb2+ on catecholamine release was studied in isolated intact and permeabilized bovine chromaffin cells. Fura-2 was used to monitor intracellular Pb2+. A characterization of Pb2(+)-fura-2 interactions in solutions simulating intracellular ionic composition showed that Pb2+ forms a 1:1 Pb2(+)-fura-2 complex (apparent dissociation constant = 4.2 x 10(-12) M, pH 7.05) whose fluorescence resembles that of the Ca2(+)-fura-2 complex. Spectra recorded from fura-2-loaded cells indicate entry of Pb2+ into the cells. Intracellular Pb2+ concentrations were proportional to extracellular Pb2+ concentrations and were found to be 10(-11)-10(-12) M in cells exposed to micromolar Pb2+ concentrations. Pb2+ elicited the release of tritiated norepinephrine from fura-2-loaded cells, indicating the extraordinary effectiveness of Pb2+ as a releasing agent. Permeabilization of cells with digitonin showed that Pb2+ is able, in the absence of Ca2+, to produce exocytotic release at concentrations of 3.2 x 10(-10) M or higher (3 orders of magnitude lower than Ca2+). These results support the notion that Pb2+ can act as a potent Ca2+ surrogate in triggering secretion.

Fast Atom Bombardment Mass Spectrometric Characterization of Poly(O-Toluidine)

1991 ◽  
Author(s):  
K. Balasaunmugam ◽  
K. G. Owens ◽  
K. F. Hsueh ◽  
P. Hoontrakul ◽  
M. A. Olsen
Keyword(s):  
Fast Atom Bombardment ◽  
Mass Spectrometric
Sign in / Sign up

Export Citation Format

Share Document