Top

Research Article

Split Viewer

Mol. Cells 2013; 36(6): 527-533

Published online December 31, 2013

https://doi.org/10.1007/s10059-013-0216-5

© The Korean Society for Molecular and Cellular Biology

The Molecular Mechanism of NELL2 Movement and Secretion in Hippocampal Progenitor HiB5 Cells

Chang Man Ha, Eun Mi Hwang, Eunju Kim, Da Yong Lee, Sunghoe Chang, Byung Ju Lee, Seong-Geun Hong, and Jae-Yong Park

1Department of Physiology, Institute of Health Science, and Medical Research Center for Neural Dysfunction, Gyeongsang National University School of Medicine, Jinju 660-290, Korea, 2Convergence Brain Research Department, Korea Brain Research Institute (KBRI), Daegu 700-010, Korea, 3Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul 136-791, Korea, 4Department of Biomedical Sciences, Neuroscience Research Institute, Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul 110-799, Korea, 5Department of Biological Sciences, University of Ulsan, Ulsan 680-749, Korea, 6These authors contributed equally to this work.

Received: July 25, 2013; Revised: November 5, 2013; Accepted: November 7, 2013

Abstract

Neural epidermal growth factor-like protein-like 2 (NELL2) is a secreted glycoprotein that is predominantly expressed in the nervous system, but little is known about the intracellular movement and secretion mechanism of this protein. By monitoring the localization and movements of enhanced green fluorescent protein (EGFP)-labeled NELL2 in living cultured hippocampal neuroprogenitor HiB5 cells, we determined the subcellular localization of NELL2 and its intracellular movement and secretion mechanism. C-terminal EGFP-fused NELL2 showed a typical expression pattern of secreted proteins, especially with respect to its localization in the endoplasmic reticulum, Golgi apparatus, and punctate structures. Vesicles containing NELL2 exhibited bidirectional movement in HiB5 cells. The majority of the vesicles (70.1%) moved in an anterograde direction with an average velocity of 0.454 ?m/s, whereas some vesicles (28.7%) showed retrograde movement with an average velocity of 0.302 ?m/s. The movement patterns of NELL2 vesicles were dependent upon the presence of microtubules in HiB5 cells. Anterograde movement of NELL2 did not lead to a detectable accumulation of NELL2 in the peripheral region of the cell, indicating that it was secreted into the culture medium. We also showed that the N-terminal 29 amino acids of NELL2 were important for secretion of this protein. Taken together, these results strongly suggest that the N-terminal region of NELL2 determines both the pattern of its intracellular expression and transport of NELL2 vesicles by high-velocity movement. Therefore, NELL2 may affect the cellular activity of cells in a paracrine or autocrine manner.

Keywords glycoprotein, intracellular movement, NELL2, secretion

Article

Research Article

Mol. Cells 2013; 36(6): 527-533

Published online December 31, 2013 https://doi.org/10.1007/s10059-013-0216-5

Copyright © The Korean Society for Molecular and Cellular Biology.

The Molecular Mechanism of NELL2 Movement and Secretion in Hippocampal Progenitor HiB5 Cells

Chang Man Ha, Eun Mi Hwang, Eunju Kim, Da Yong Lee, Sunghoe Chang, Byung Ju Lee, Seong-Geun Hong, and Jae-Yong Park

1Department of Physiology, Institute of Health Science, and Medical Research Center for Neural Dysfunction, Gyeongsang National University School of Medicine, Jinju 660-290, Korea, 2Convergence Brain Research Department, Korea Brain Research Institute (KBRI), Daegu 700-010, Korea, 3Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul 136-791, Korea, 4Department of Biomedical Sciences, Neuroscience Research Institute, Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul 110-799, Korea, 5Department of Biological Sciences, University of Ulsan, Ulsan 680-749, Korea, 6These authors contributed equally to this work.

Received: July 25, 2013; Revised: November 5, 2013; Accepted: November 7, 2013

Abstract

Neural epidermal growth factor-like protein-like 2 (NELL2) is a secreted glycoprotein that is predominantly expressed in the nervous system, but little is known about the intracellular movement and secretion mechanism of this protein. By monitoring the localization and movements of enhanced green fluorescent protein (EGFP)-labeled NELL2 in living cultured hippocampal neuroprogenitor HiB5 cells, we determined the subcellular localization of NELL2 and its intracellular movement and secretion mechanism. C-terminal EGFP-fused NELL2 showed a typical expression pattern of secreted proteins, especially with respect to its localization in the endoplasmic reticulum, Golgi apparatus, and punctate structures. Vesicles containing NELL2 exhibited bidirectional movement in HiB5 cells. The majority of the vesicles (70.1%) moved in an anterograde direction with an average velocity of 0.454 ?m/s, whereas some vesicles (28.7%) showed retrograde movement with an average velocity of 0.302 ?m/s. The movement patterns of NELL2 vesicles were dependent upon the presence of microtubules in HiB5 cells. Anterograde movement of NELL2 did not lead to a detectable accumulation of NELL2 in the peripheral region of the cell, indicating that it was secreted into the culture medium. We also showed that the N-terminal 29 amino acids of NELL2 were important for secretion of this protein. Taken together, these results strongly suggest that the N-terminal region of NELL2 determines both the pattern of its intracellular expression and transport of NELL2 vesicles by high-velocity movement. Therefore, NELL2 may affect the cellular activity of cells in a paracrine or autocrine manner.

Keywords: glycoprotein, intracellular movement, NELL2, secretion

Mol. Cells
Aug 31, 2022 Vol.45 No.8, pp. 513~602
COVER PICTURE
Cryo-EM structure of human porphyrin transporter ABCB6 (main figure) shows that binding of hemin (inset, magenta) in concert with two glutathione molecules (cyan) primes ABCB6 for high ATP turnover (Kim et al., pp. 575-587).

Share this article on

  • line
  • mail

Related articles in Mol. Cells

Molecules and Cells

eISSN 0219-1032
qr-code Download