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Title: Myristoylation restricts orientation of the GRASP domain on membranes and promotes membrane tethering
Authors: Heinrich, Frank
Nanda, Hirsh
Goh, Haw Zan
Bachert, Collin
Lösche, Mathias
Linstedt, Adam D.
Keywords: DRNTU::Engineering::Materials
Issue Date: 2014
Source: Heinrich, F., Nanda, H., Goh, H. Z., Bachert, C., Lösche, M., & Linstedt, A. D. (2014). Myristoylation restricts orientation of the GRASP domain on membranes and promotes membrane tethering. Journal of Biological Chemistry, 289(14), 9683-9691.
Series/Report no.: Journal of Biological Chemistry
Abstract: The mammalian Golgi reassembly stacking protein (GRASP) proteins are Golgi-localized homotypic membrane tethers that organize Golgi stacks into a long, contiguous ribbon-like structure. It is unknown how GRASPs undergo trans pairing given that cis interactions between the proteins in the plane of the membrane are intrinsically favored. To test the hypothesis that myristoylation of the self-interacting GRASP domain restricts its orientation on the membrane to favor trans pairing, we established an in vitro assay that recapitulates GRASP-dependent membrane tethering and used neutron reflection under similar conditions to determine the orientation of the GRASP domain. In vivo, the membrane association of GRASP proteins is conferred by the simultaneous insertion of an N-terminal myristic acid and binding to a Golgi-associated binding partner. In our assay, the latter contact was replaced using a C-terminal hexa-His moiety, which bound to Ni2+-conjugated lipids incorporated into a substrate-supported bilayer lipid membrane. Nonmyristoylated protein lacked a fixed orientation on the membrane and inefficiently tethered liposomes. In contrast, myristoylated GRASP promoted tethering and exhibited a unique membrane complex. Thus, myristoylation restricts the membrane orientation of the GRASP domain favoring interactions in trans for membrane tethering.
ISSN: 0021-9258
Rights: © 2014 The American Society for Biochemistry and Molecular Biology, Inc. This paper was published in Journal of Biological Chemistry and is made available as an electronic reprint (preprint) with permission of The American Society for Biochemistry and Molecular Biology, Inc. The published version is available at: []. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.
Fulltext Permission: open
Fulltext Availability: With Fulltext
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