Banks Music Hall in Lendal ceased trading yesterday.
The music store used to stock musical instruments, sheet music and vinyl records.
According to York Explore Library and Archive, Banks Music Room is believed to be the oldest music shop in the UK.
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It first opened on Blake Street in 1756 under the ownership of Thomas Haxby, before changing hands and moving to various locations in the city centre.
The store has been in its current location in Lendal since 1985.
The Musicroom was part of the Hal Leonard Group, which confirmed the closure last month as part of plans to restructure its UK and Ireland business.