Phosphorylation and O-Glycosylation Sites of Human Chromogranin A (CGA79–439) from Urine of Patients with Carcinoid Tumors

Bühling

The Journal of Immunology

et al. 2001

Human cathepsin W (also called lymphopain) is a recently described papain-like cysteine protease of unknown function whose gene expression was found to be restricted to cytotoxic cells. Here we demonstrate that cathepsin W is expressed predominantly in NK cells and, to a lesser extent, in CTLs. Quantitative RT-PCR revealed that NK cells contained ∼21 times more cathepsin W transcript than CTLs. The predominant expression of cathepsin W in NK cells was further confirmed by Western blot analysis and immunohistochemistry. IL-2-mediated stimulation of NK cells and CTLs revealed a stronger up-regulation of the cathepsin W gene and protein expression in NK cells (7-fold) than in CTLs (2-fold). Transfection experiments of HeLa cells and biochemical analyses revealed that cathepsin W is exclusively “high mannose-type” glycosylated and is mainly targeted to the endoplasmic reticulum (ER). Interestingly, the ER localization of cathepsin W was also found in NK cells, in which colocalization studies revealed an overlapping staining of cathepsin W and Con A, an ER-specific lectin. Furthermore, subcellular fractionation of cathepsin W-expressing cells confirmed the ER localization and showed that cathepsin W is membrane associated. Based on the results of this study, cathepsin W might represent a putative component of the ER-resident proteolytic machinery. The constitutive expression in NK cells and the stronger up-regulation of cathepsin W by IL-2 in NK cells than CTLs suggest that cathepsin W is not just a marker of cytotoxic cells but is, rather, specifically expressed in NK cells.

Section: Biochemical Characterization Of Cathepsin Wmentioning

confidence: 99%

Human Cathepsin W, a Cysteine Protease Predominantly Expressed in NK Cells, Is Mainly Localized in the Endoplasmic Reticulum

Bühling

The Journal of Immunology

et al. 2001

Journal of Biological Chemistry

“…Absorbance was monitored at 214 nm, and elution was performed with a linear gradient as indicated on the right-hand scale. Arrow indicates the fraction corresponding to the antifungal peptide CGA [47][48][49][50][51][52][53][54][55][56][57][58][59][60] .…”

Section: Identification Of Antifungal Rhvs-1-derived Peptidesmentioning

confidence: 99%

“…2), we decided to examine the role of the tripeptide Arg 43 -Gly 44 -Asp 45 and the length of the peptidic chain, testing the activities of the synthetic peptides CGA [41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56][57][58][59][60] , CGA , and CGA , respectively (Table III). The comparison of their activity with the activity of CGA [47][48][49][50][51][52][53][54][55][56][57][58][59][60] highlights several important points as follows: (i) CGA 41 ments CGA [47][48][49][50][51][52][53][54][55][56]…”

Section: Identification Of Antifungal Rhvs-1-derived Peptidesmentioning

confidence: 99%

“…Then, the antifungal properties of the peptides included in the different peaks were tested. An active peptide corresponding to CGA [47][48][49][50][51][52][53][54][55][56][57][58][59][60] with an experimental molecular mass of 1732.9 Da ( Fig. 3B; expected molecular mass, 1733.1 Da) is active against N. crassa showing a minimal inhibitory concentration giving 100% inhibition of 7 M. Incubation of fungi with synthetic peptide corresponding to CGA [47][48][49][50][51][52][53][54][55][56][57][58][59][60] at increasing concentrations from 3 to 10 M (Table III) induces a decrease of fungal proliferation and the inhibition in the growth of filaments.…”

Section: Identification Of Antifungal Rhvs-1-derived Peptidesmentioning

confidence: 99%

See 1 more Smart Citation

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

Lugardon¹,

Raffner²,

Goumon³

et al. 2000

Self Cite

151

140

Vasostatin-1, the natural N-terminal 1-76 chromogranin A (CGA)-derived fragment in bovine sequence, has been purified from chromaffin secretory granules and identified by sequencing and matrix-assisted laser desorption time-of-flight mass spectrometry. This peptide, which displays antibacterial activity against Gram-positive bacteria at micromolar concentrations, is also able to kill a large variety of filamentous fungi and yeast cells in the 1-10 M range. We have found that the C-terminal moiety of vasostatin-1 is essential for the antifungal activity, and shorter active peptides have been synthesized. In addition, from the comparison with the activity displayed by related peptides (human recombinant and rat synthetic fragments), we could determine that antibacterial and antifungal activities have different structural requirements. To assess for such activities in vivo, CGA and CGA-derived fragments were identified in secretory material released from human polymorphonuclear neutrophils upon stimulation. Vasostatin-1, which is stored in a large variety of cells (endocrine, neuroendocrine, and neurons) and which is liberated from stimulated chromaffin and immune cells upon stress, may represent a new component active in innate immunity.

Results and Problems in Cell Differentiation

“…As illustrated in Fig. 1a, prochromacin is the largest VS-I free CgA peptide in bovine chromaffin granules (Metz-Boutigue et al 1993; and occurs also as the main CgA productin the urine of carcinoid patients (Gadroy et al 1998).…”

Section: Cgamentioning

confidence: 99%

Chromogranins A and B and Secretogranin II as Prohormones for Regulatory Peptides from the Diffuse Neuroendocrine System

Helle

2010

Chromogranin A (CgA), chromogranin B (CgB), and secretogranin II (SgII) belong to a family of uniquely acidic secretory proteins in elements of the diffuse neuroendocrine system. These "granins" are characterized by numerous pairs of basic amino acids as potential sites for intra- and extragranular processing. In response to adequate stimuli, the granins are coreleased with neurotransmitters and hormones and appear in the circulation as potential modulators of homeostatic processes. This review is directed towards functional aspects of the secreted CgA, CgB, and SgII and their biologically active sequences. Widely different effects and targets have been reported for granin-derived peptides. So far, the CgA peptides vasostatin-I, pancreastatin, and catestatin, the CgB peptides CgB(1-41) and secretolytin, and the SgII peptide secretoneurin are the most likely candidates for granin-derived regulatory peptides. Most of their effects fit into patterns of direct or indirect modulations of major functions, in particular associated with inflammatory conditions.