Functional role of the interaction between polysialic acid and myristoylated alanine-rich C kinase substrate at the plasma membrane.
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Functional role of the interaction between polysialic acid and myristoylated alanine-rich C kinase substrate at the plasma membrane. / Theis, Thomas; Mishra, Bibhudatta; von der Ohe, Maren; Loers, Gabriele; Prondzynski, Maksymilian; Pless, Ole; Blackshear, Perry J; Schachner, Melitta; Kleene, Ralf.
In: J BIOL CHEM, Vol. 288, No. 9, 9, 2013, p. 6726-6742.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Functional role of the interaction between polysialic acid and myristoylated alanine-rich C kinase substrate at the plasma membrane.
AU - Theis, Thomas
AU - Mishra, Bibhudatta
AU - von der Ohe, Maren
AU - Loers, Gabriele
AU - Prondzynski, Maksymilian
AU - Pless, Ole
AU - Blackshear, Perry J
AU - Schachner, Melitta
AU - Kleene, Ralf
PY - 2013
Y1 - 2013
N2 - Polysialic acid (PSA) is a homopolymeric glycan that plays crucial roles in the developing and adult nervous system. So far only a few PSA-binding proteins have been identified. Here, we identify myristoylated alanine-rich C kinase substrate (MARCKS) as novel PSA binding partner. Binding assays showed a direct interaction between PSA and a peptide comprising the effector domain of MARCKS (MARCKS-ED). Co-immunoprecipitation of PSA-carrying neural cell adhesion molecule (PSA-NCAM) with MARCKS and co-immunostaining of MARCKS and PSA at the cell membrane of hippocampal neurons confirm the interaction between PSA and MARCKS. Co-localization and an intimate interaction of PSA and MARCKS at the cell surface was seen by confocal microscopy and fluorescence resonance energy transfer (FRET) analysis after the addition of fluorescently labeled PSA or PSA-NCAM to live CHO cells or hippocampal neurons expressing MARCKS as a fusion protein with green fluorescent protein (GFP). Cross-linking experiments showed that extracellularly applied PSA or PSA-NCAM and intracellularly expressed MARCKS-GFP are in close contact, suggesting that PSA and MARCKS interact with each other at the plasma membrane from opposite sides. Insertion of PSA and MARCKS-ED peptide into lipid bilayers from opposite sides alters the electric properties of the bilayer confirming the notion that PSA and the effector domain of MARCKS interact at and/or within the plane of the membrane. The MARCKS-ED peptide abolished PSA-induced enhancement of neurite outgrowth from cultured hippocampal neurons indicating an important functional role for the interaction between MARCKS and PSA in the developing and adult nervous system.
AB - Polysialic acid (PSA) is a homopolymeric glycan that plays crucial roles in the developing and adult nervous system. So far only a few PSA-binding proteins have been identified. Here, we identify myristoylated alanine-rich C kinase substrate (MARCKS) as novel PSA binding partner. Binding assays showed a direct interaction between PSA and a peptide comprising the effector domain of MARCKS (MARCKS-ED). Co-immunoprecipitation of PSA-carrying neural cell adhesion molecule (PSA-NCAM) with MARCKS and co-immunostaining of MARCKS and PSA at the cell membrane of hippocampal neurons confirm the interaction between PSA and MARCKS. Co-localization and an intimate interaction of PSA and MARCKS at the cell surface was seen by confocal microscopy and fluorescence resonance energy transfer (FRET) analysis after the addition of fluorescently labeled PSA or PSA-NCAM to live CHO cells or hippocampal neurons expressing MARCKS as a fusion protein with green fluorescent protein (GFP). Cross-linking experiments showed that extracellularly applied PSA or PSA-NCAM and intracellularly expressed MARCKS-GFP are in close contact, suggesting that PSA and MARCKS interact with each other at the plasma membrane from opposite sides. Insertion of PSA and MARCKS-ED peptide into lipid bilayers from opposite sides alters the electric properties of the bilayer confirming the notion that PSA and the effector domain of MARCKS interact at and/or within the plane of the membrane. The MARCKS-ED peptide abolished PSA-induced enhancement of neurite outgrowth from cultured hippocampal neurons indicating an important functional role for the interaction between MARCKS and PSA in the developing and adult nervous system.
KW - Animals
KW - Mice
KW - Mice, Knockout
KW - CHO Cells
KW - Cricetinae
KW - Cricetulus
KW - Nerve Tissue Proteins/genetics/metabolism
KW - Membrane Proteins/genetics/metabolism
KW - Hippocampus/cytology/metabolism
KW - Cell Membrane/genetics/metabolism
KW - Neural Cell Adhesion Molecule L1/genetics/metabolism
KW - Intracellular Signaling Peptides and Proteins/genetics/metabolism
KW - Lipid Bilayers
KW - Neurites/metabolism
KW - Peptides/genetics/metabolism/pharmacology
KW - Sialic Acids/genetics/metabolism
KW - Animals
KW - Mice
KW - Mice, Knockout
KW - CHO Cells
KW - Cricetinae
KW - Cricetulus
KW - Nerve Tissue Proteins/genetics/metabolism
KW - Membrane Proteins/genetics/metabolism
KW - Hippocampus/cytology/metabolism
KW - Cell Membrane/genetics/metabolism
KW - Neural Cell Adhesion Molecule L1/genetics/metabolism
KW - Intracellular Signaling Peptides and Proteins/genetics/metabolism
KW - Lipid Bilayers
KW - Neurites/metabolism
KW - Peptides/genetics/metabolism/pharmacology
KW - Sialic Acids/genetics/metabolism
M3 - SCORING: Journal article
VL - 288
SP - 6726
EP - 6742
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 9
M1 - 9
ER -
