Publikationsdatenbank

Functional characterization of palmitoylated and nonacylated SNAP-25 purified from insect cells infected with recombinant baculovirus (2003)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Kammer, Bettina

Schmidt, Michael F. G.

Veit, Michael (WE 5)

Quelle

Molecular and cellular neuroscience : MCN

Bandzählung: 23

Heftzählung: 3

Seiten: 333 – 340

ISSN: 1044-7431

Sprache

Englisch

Verweise

URL (Volltext): https://pubmed.ncbi.nlm.nih.gov/12837618/

DOI: 10.1016/s1044-7431(03)00064-2

Pubmed: 12837618

Kontakt

Institut für Virologie

Robert-von-Ostertag-Str. 7-13
14163 Berlin
+49 30 838 51833
virologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

SNARE complex formation among syntaxin 1, VAMP 2, and SNAP-25 triggers fusion of synaptic vesicles with the presynaptic plasma membrane. After fusion the SNARE complex is disassembled by NSF and alpha-SNAP. These reactions have already been characterized with recombinant proteins lacking the authentic protein modifications. To study the role of palmitoylation of SNAP-25, we have purified 6xHis-SNAP-25, the wild-type protein, and mutants with deletions in the palmitoylation region from insect cells infected with recombinant baculovirus. Metabolic labeling with [(3)H]palmitate and Triton-X-114 phase distribution proved that SNAP-25 is palmitoylated in significant amounts. Palmitoylated and nonacylated SNAP-25 form SDS-resistant SNARE complexes which could then be disassembled by NSF and alpha-SNAP. Palmitoylated SNAP-25 attaches almost quantitatively to liposomes, whereas only small amounts of nonacylated SNAP-25 bind to these artificial membranes. Thus, palmitoylation of SNAP-25 is not required for assembly and disassembly of the SNARE complex, but for stable membrane anchoring of this intrinsically hydrophilic protein.