Regulation of the neuronal SNARE-complex by accessory proteins

Regulation of the neuronal SNARE-complex by accessory proteins

Shrutee Jakhanwal

Doctoral thesis

Date of Examination: 2017-07-13

Date of issue: 2018-02-16

Advisor: Jahn, Reinhard Prof. Dr.

Referee: Steinem, Claudia Prof. Dr.

Referee: Rizzoli, Silvio Prof. Dr.

Persistent Address: http://hdl.handle.net/11858/00-1735-0000-002E-E355-D

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Abstract

English

In this work, I have thoroughly characterized a novel complex containing syntaxin1a (1-288), SNAP25a and Munc18-1 and established it as an efficient acceptor complex for synaptobrevin-binding, using several biochemical and biophysical techniques. The use of all full-length proteins provide tremendous strength to the study, giving it a ‘near-native’ scenario for the behaviors of the proteins being discussed. In addition to understanding the functional importance of this complex, I also attained its structural overview using chemical crosslinking assays. And, finally, by testing the resistance of this complex to disassembly by NSF-αSNAP, I could establish that the syntaxin1a/SNAP25a/Munc18-1 complex can allow SNARE-assembly to proceed in an NSF-αSNAP-resistant manner.

Keywords: SNAREs, Munc18, synaptic vesicle exocytosis