The Substrates of a Sodium- and Chloride-Coupled γ-Aminobutyric Acid Transporter Protect Multiple Sites throughout the Protein against Proteolytic Cleavage

Nicola J. Mabjeesh, Baruch I. Kanner

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Fragments of the (Na+ + Cl)-coupled GABAA transporter were produced by proteolysis of membrane vesicles and reconstituted preparations from rat brain. The former were digested with Pronase, the latter with trypsin. Fragments with different apparent molecular masses were recognized by sequence-directed antibodies raised against this transporter. When GABA was present in the digestion medium, the generation of these fragments was almost entirely blocked. At the same time, the neurotransmitter largely prevented the loss of activity caused by the protease. The effect was specific for GABA; protection was not afforded by other neurotransmitters. It was only observed when the two cosubstrates, sodium and chloride, were present on the same side of the membrane as GABA. The results indicate that the transporter may exist in two conformations. In the absence of one or more of the substrates, multiple sites located throughout the transporter are accessible to the proteases. In the presence of all three substrates—conditions favoring the formation of the translocation complex—the conformation is changed such that these sites become inaccessible to protease action.

Original languageEnglish
Pages (from-to)8540-8546
Number of pages7
JournalBiochemistry
Volume32
Issue number33
DOIs
StatePublished - 1 Jan 1993
Externally publishedYes

Fingerprint

Dive into the research topics of 'The Substrates of a Sodium- and Chloride-Coupled γ-Aminobutyric Acid Transporter Protect Multiple Sites throughout the Protein against Proteolytic Cleavage'. Together they form a unique fingerprint.

Cite this