Skip to content

Research at St Andrews

Protein kinase B/Akt is a novel cysteine string protein kinase that regulates exocytosis release kinetics and quantal size

Research output: Contribution to journalArticlepeer-review

DOI

Author(s)

G J O Evans, J W Barclay, G R Prescott, S R Jo, R D Burgoyne, M J Birnbaum, A Morgan

School/Research organisations

Abstract

Protein kinase B/Akt has been implicated in the insulin-dependent exocytosis of GLUT4-containing vesicles, and, more recently, insulin secretion. To determine if Akt also regulates insulin-independent exocytosis, we used adrenal chromaffin cells, a popular neuronal model. Akt1 was the predominant isoform expressed in chromaffin cells, although lower levels of Akt2 and Akt3 were also found. Secretory stimuli in both intact and permeabilized cells induced Akt phosphorylation on serine 473, and the time course of Ca2+-induced Akt phosphorylation was similar to that of exocytosis in permeabilized cells. To determine if Akt modulated exocytosis, we transfected chromaffin cells with Akt constructs and monitored catecholamine release by amperometry. Wild-type Akt had no effect on the overall number of exocytotic events, but slowed the kinetics of catecholamine release from individual vesicles, resulting in an increased quantal size. This effect was due to phosphorylation by Akt, because it was not seen in cells transfected with kinase-dead mutant Akt. As overexpression of cysteine string protein (CSP) results in a similar alteration in release kinetics and quantal size, we determined if CSP was an Akt substrate. In vitro P-32-phosphorylation studies revealed that Akt phosphorylates CSP on serine 10. Using phospho-Ser10-specific antisera, we found that both transfected and endogenous cellular CSP is phosphorylated by Akt on this residue. Taken together, these findings reveal a novel role for Akt phosphorylation in regulating the late stages of exocytosis and suggest that this is achieved via the phosphorylation of CSP on serine 10.

Close

Details

Original languageEnglish
Pages (from-to)1564-1572
Number of pages9
JournalJournal of Biological Chemistry
Volume281
Issue number3
DOIs
Publication statusPublished - 20 Jan 2006

    Research areas

  • ADRENAL CHROMAFFIN CELLS, STIMULATED GLUT4 TRANSLOCATION, SYNAPTIC VESICLE PROTEIN, GRANULE EXOCYTOSIS, 3T3-L1 ADIPOCYTES, GLUCOSE-UPTAKE, DEPENDENT PHOSPHORYLATION, WORTMANNIN INHIBITION, INSULIN STIMULATION, TRANSMITTER RELEASE

Discover related content
Find related publications, people, projects and more using interactive charts.

View graph of relations

Related by author

  1. Phosphorylation of Cysteine string protein triggers a major conformational switch

    Patel, P., Prescott, G. R., Burgoyne, R. D., Lian, L-Y. & Morgan, A., 2 Aug 2016, In: Structure. 24, 8, p. 1380-1386 7 p.

    Research output: Contribution to journalArticlepeer-review

  2. Endoplasmic Reticulum Localization of DHHC Palmitoyltransferases Mediated by Lysine-based Sorting Signals

    Gorleku, O. A., Barns, A-M., Prescott, G. R., Greaves, J. & Chamberlain, L. H., 11 Nov 2011, In: Journal of Biological Chemistry. 286, 45, p. 39573-39584 12 p.

    Research output: Contribution to journalArticlepeer-review

  3. Regional and developmental brain expression patterns of SNAP25 splice variants

    Prescott, G. R. & Chamberlain, L. H., 28 Apr 2011, In: BioMed Central Neuroscience. 12, 9 p., 35.

    Research output: Contribution to journalArticlepeer-review

  4. Structure-function study of mammalian Munc18-1 and C. elegans UNC-18 implicates domain 3b in the regulation of exocytosis

    Graham, M. E., Prescott, G. R., Johnson, J. R., Jones, M., Walmesley, A., Haynes, L. P., Morgan, A., Burgoyne, R. D. & Barclay, J. W., 21 Mar 2011, In: PLoS One. 6, 3, 12 p., e17999.

    Research output: Contribution to journalArticlepeer-review

  5. Regulation of SNAP-25 trafficking and function by palmitoylation

    Greaves, J., Prescott, G. R., Gorleku, O. A. & Chamberlain, L. H., Feb 2010, In: Biochemical Society Transactions. 38, p. 163-166 4 p.

    Research output: Contribution to journalArticlepeer-review

Related by journal

  1. Journal of Biological Chemistry (Journal)

    Simon John Powis (Member of editorial board)

    1 Jun 2015

    Activity: Publication peer-review and editorial work typesEditor of research journal

  2. Journal of Biological Chemistry (Journal)

    Terry K Smith (Member of editorial board)

    2006 → …

    Activity: Publication peer-review and editorial work typesPeer review of manuscripts

Related by journal

  1. Iron is a ligand of SecA-like metal-binding domains in vivo

    Cranford-Smith, T., Jamshad, M., Jeeves, M., Chandler, R. A., Yule, J., Robinson, A., Alam, F., Dunne, K. A., Aponte Angarita, E. H., Alanazi, M., Carter, C., Henderson, I. R., Lovett, J. E., Winn, P., Knowles, T. & Huber, D., 2 Apr 2020, In: Journal of Biological Chemistry. Papers in Press

    Research output: Contribution to journalArticlepeer-review

  2. The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

    Koo, C. Z., Harrison, N., Noy, P. J., Szyroka, J., Matthews, A. L., Hsia, H-E., Müller, S. A., Tüshaus, J., Goulding, J., Willis, K., Apicella, C., Cragoe, B., Davis, E., Keles, M., Malinova, A., McFarlane, T. A., Morrison, P. R., Nguyen, H. T. H., Sykes, M. C., Ahmed, H. & 11 others, Di Maio, A., Seipold, L., Saftig, P., Cull, E., Pliotas, C., Rubinstein, E., Poulter, N. S., Briddon, S. J., Holliday, N. D., Lichtenthaler, S. F. & Tomlinson, M. G., 28 Feb 2020, In: Journal of Biological Chemistry.

    Research output: Contribution to journalArticlepeer-review

  3. Combining SPR with atomic-force microscopy enables singlemolecule insights into activation and suppression of the complement cascade

    Makou, E., Bailey, R. G., Johnston, H., Parkin, J. D., Hulme, A. N., Hähner, G. & Barlow, P. N., 27 Dec 2019, In: Journal of Biological Chemistry. 294, 52, p. 20148-20163 16 p.

    Research output: Contribution to journalArticlepeer-review

  4. Dysregulated Zn2+ homeostasis impairs cardiac type-2 ryanodine receptor and mitsugumin 23 functions, leading to sarcoplasmic reticulum Ca2+ leakage

    Reilly-O'Donnell, B., Robertson, G. B., Karumbi, A., McIntyre, C., Bal, W., Nishi, M., Takeshima, H., Stewart, A. J. & Pitt, S. J., 11 Aug 2017, In: Journal of Biological Chemistry. 292, 32, p. 13361-13373 13 p.

    Research output: Contribution to journalArticlepeer-review

  5. The major histocompatibility complex class I immunopeptidome of extracellular vesicles

    Synowsky, S. A., Shirran, S. L., Cooke, F. G. M., Antoniou, A. N., Botting, C. H. & Powis, S. J., 13 Oct 2017, In: Journal of Biological Chemistry. 292, p. 17084-17092

    Research output: Contribution to journalArticlepeer-review

ID: 7221416

Top