Sorting of the Neuroendocrine Secretory Protein Secretogranin II into the Regulated Secretory Pathway

Courel, Maïté; Vasquez, Michael S.; Hook, Vivian; Mahata, Sushil K.; Taupenot, Laurent

doi:10.1074/jbc.m709832200

Cited by 50 publications

(48 citation statements)

References 53 publications

(170 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since both helices interact with the micelles through their hydrophobic face and we did not observe interactions between the two helices, it appears that these structural elements interact with the micelle (and the membrane) in a carpet-like mechanism (14), effectively laying on the membrane surface. Interestingly, two similar DCSG-sorting motifs have also been observed in SgII (15). SgII contains both an amino-terminal and a carboxy-terminal sorting domain, and both of these domains have been predicted to form helix-loop-helix motifs.…”

Section: Discussionmentioning

confidence: 86%

“…Mouse PC1/3 cDNA (NM013628) encoding amino acids 711-753 was inserted in to the pGEX-4T (GE Healthcare) vector and expressed in E.Coli Topp2 cells. Cells were grown in 2 L of LB or minimal media containing 15 NH 4 Cl and/or 13 C-glucose and induced with IPTG for 3 h at 30°C. Pellets were resuspended in 100 ml of Buffer A (20 mM Tris, pH ϭ 7.4, 1 M NaCl, 1 mM EDTA and 1 mM DTT) and lysed.…”

Section: Methodsmentioning

confidence: 99%

“…S2). The addition of 20 mM CHAPS or 200 mM DPC to PC1/3 711-753 at NMR concentrations allowed for the complete 1 H, 15 N and 13 C chemical shift assignment. The use of micelle-forming conditions such as CHAPS and DPC micelles is consistent with previous reports that PC1/3 associates with membranes both in transfected cells (7) and in vitro (8).…”

Section: Dcsg-sorting Domain Of Pc1/3 In a Micellarmentioning

confidence: 99%

“…The interproton distances were estimated from the intensities of the cross peaks observed in the 3D 15 N-edited NOESY-HSQC (30, 31) and 13 C-edited heteronuclear multiple quantum coherence-total correlation (HMQC)-NOESY experiments (32). The backbone dihedral angles were obtained from the analysis of 1 H, 15 N, 13 C', 13 C ␣ , and 13 C ␤ chemical shifts using the program TALOS (33). Structure calculations were performed with the program CNS (34) using a combination of torsion angle dynamics (TAD) and Cartesian dynamics.…”

Section: Nmr Sample Preparation Andmentioning

confidence: 99%

See 3 more Smart Citations

Functional and structural characterization of a dense core secretory granule sorting domain from the PC1/3 protease

Dikeakos

Lello

Lacombe

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Several peptide hormones are initially synthesized as inactive precursors. It is only on entry of these prohormones and their processing proteases into dense core secretory granules (DCSGs) that the precursors are cleaved to generate their active forms. Prohormone convertase (PC)1/3 is a processing protease that is targeted to DCSGs. The signal for targeting PC1/3 to DCSGs resides in its carboxy-terminal tail (PC1/3 617-753), where 3 regions (PC1/ 3 617-625, PC1/3665-682, and PC1/3711-753) are known to aid in sorting and membrane association. In this article, we have determined a high-resolution structure of the extreme carboxy-terminal sorting domain, PC1/3 711-753 in micelles by NMR spectroscopy. PC1/3711-753 contains 2 alpha helices located between residues 722-728 and 738 -750. Functional assays demonstrate that the second helix (PC1/3 738 -750) is necessary and sufficient to target a constitutively secreted protein to granules, and that L 745 anchors a hydrophobic patch that is critical for sorting. Also, we demonstrate that calcium binding by the second helix of PC1/3 711-753 promotes aggregation of the domain via the hydrophobic patch centered on L 745 . These results provide a structure-function analysis of a DCSG-sorting domain, and reveal the importance of a hydrophobic patch and calcium binding in controlling the sorting of proteins containing alpha helices to DCSGs.NMR ͉ prohormone convertases D ense core secretory granules (DCSGs) are a repository of proteases and hormones in endocrine and neuroendocrine cells. Many of these proteins are first synthesized as precursors that are activated by protease cleavage after the precursor enters the nascent secretory granules (1-3). The processed proteins are stored within the DCSGs and released when the cell receives the correct stimulus (4). This regulated secretory pathway differs from the pathway of constitutive protein secretion, where proteins are packaged in low-density vesicles and released in an unregulated manner. Thus, the targeting of the protein precursors and their processing enzymes to DCSGs is critical to ensure the secretion of protein hormones such as insulin, glucagon and proteases such as renin.Two models have been postulated to explain protein sorting to DCSGs (5). The 'sorting by entry' model postulates that proteins are triaged at the trans Golgi network (TGN) via a proteinreceptor interaction and proteins interacting with this receptor are diverted into the regulated pathway. A second model is 'sorting by retention' where all proteins, regardless of their final destination, enter an immature secretory granule from which proteins destined for constitutive secretion or targeted to other organelles would be extruded while proteins destined to be stored in the mature DCSG for regulated release would be selectively retained. Nascent granules mature through a process that involves immature granule fusion, acidification, increased calcium content and prohormone processing and condensation (1). Regardless of how DCSG-resident proteins are selec...

show abstract

Section: Discussionmentioning

confidence: 86%

Section: Methodsmentioning

confidence: 99%

Section: Dcsg-sorting Domain Of Pc1/3 In a Micellarmentioning

confidence: 99%

Section: Nmr Sample Preparation Andmentioning

confidence: 99%

See 2 more Smart Citations

Functional and structural characterization of a dense core secretory granule sorting domain from the PC1/3 protease

Dikeakos

Lello

Lacombe

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…SCG2 is able to induce the formation of secretory granule-like structures in nonneuroendocrine cells, indicating a crucial role of SCG2 in generating secretory vesicles, and functions in the packaging and sorting of peptide hormones and neuropeptides into secretory vesicles (5). Two domains on the SCG2 full-length protein (domain spanning human SCG2 residues 25 to 41 [hSCG2 [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41] ] and hSCG2 334 -348 ) are target-ing signals for sorting of the protein into regulated secretory pathway granules (6). The full-length mammalian SCG2 protein contains 617 residues and is cleaved to generate three bioactive peptides: secretoneurin (SN) (rat SgII 154 -186 ), EM66 (rat SgII 189 -256 ), and manserin (rat SgII 529 -568 ) (4,7).…”

mentioning

confidence: 99%

Differential Reovirus-Specific and Herpesvirus-Specific Activator Protein 1 Activation of Secretogranin II Leads to Altered Virus Secretion

Berard

Severini

Coombs

2015

J Virol

View full text Add to dashboard Cite

Viruses utilize host cell machinery for propagation and manage to evade cellular host defense mechanisms in the process. Much remains unknown regarding how the host responds to viral infection. We recently performed global proteomic screens of mammalian reovirus TIL-and T3D-infected and herpesvirus (herpes simplex virus 1 [HSV-1])-infected HEK293 cells. The nonenveloped RNA reoviruses caused an upregulation, whereas the enveloped DNA HSV-1 caused a downregulation, of cellular secretogranin II (SCG2). SCG2, a member of the granin family that functions in hormonal peptide sorting into secretory vesicles, has not been linked to virus infections previously. We confirmed SCG2 upregulation and found SCG2 phosphorylation by 18 h postinfection (hpi) in reovirus-infected cells. We also found a decrease in the amount of reovirus secretion from SCG2 knockdown cells. Similar analyses of cells infected with HSV-1 showed an increase in the amount of secreted virus. Analysis of the stress-activated protein kinase (SAPK)/Jun N-terminal protein kinase (JNK) pathway indicated that each virus activates different pathways leading to activator protein 1 (AP-1) activation, which is the known SCG2 transcription activator. We conclude from these experiments that the negative correlation between SCG2 quantity and virus secretion for both viruses indicates a virusspecific role for SCG2 during infection. IMPORTANCEMammalian reoviruses affect the gastrointestinal system or cause respiratory infections in humans. Recent work has shown that all mammalian reovirus strains (most specifically T3D) may be useful oncolytic agents. The ubiquitous herpes simplex viruses cause common sores in mucosal areas of their host and have coevolved with hosts over many years. Both of these virus species are prototypical representatives of their viral families, and investigation of these viruses can lead to further knowledge of how they and the other more pathogenic members of their respective families interact with the host. Here we show that secretogranin II (SCG2), a protein not previously studied in the context of virus infections, alters virus output in a virus-specific manner and that the quantity of SCG2 is inversely related to amounts of infectious-virus secretion. Herpesviruses may target this protein to facilitate enhanced virus release from the host. Viruses that infect a host cell rely on the host machinery to replicate while managing to evade the immune system. Thus, there is an intricate relationship between the host and the virus that can be characterized by measuring changes in the host's protein effector molecules during the course of infection. In a previous study (1), we identified numerous HEK293 protein abundance changes after infection with the prototypic mammalian reovirus subtype 1 Lang (T1L), including the upregulation of secretogranin II (SCG2) at 24 h postinfection (hpi).SCG2 is a protein in the structurally and functionally related granin family that also includes chromogranins A and B, secretogranin III, secretogranin V (se...

show abstract

Secretogranin II: Novel Insights into Expression and Function of the Precursor of the Neuropeptide Secretoneurin

Fischer‐Colbrie

Theurl

Kirchmair

2017

Chromogranins: From Cell Biology to Physiology and Biomedicine

View full text Add to dashboard Cite

Sorting of the Neuroendocrine Secretory Protein Secretogranin II into the Regulated Secretory Pathway

Cited by 50 publications

References 53 publications

Functional and structural characterization of a dense core secretory granule sorting domain from the PC1/3 protease

Functional and structural characterization of a dense core secretory granule sorting domain from the PC1/3 protease

Differential Reovirus-Specific and Herpesvirus-Specific Activator Protein 1 Activation of Secretogranin II Leads to Altered Virus Secretion

Secretogranin II: Novel Insights into Expression and Function of the Precursor of the Neuropeptide Secretoneurin

Contact Info

Product

Resources

About