Oxfam Australia's Launceston store will close alongside all of the charity's retail, wholesale and online trading.
Oxfam Australia chief operating officer Tony McKimmie said closing was a difficult decision, but it was made in the face of ongoing commercial pressures and a tough retail environment.
Two employees, one permanent and the other part-time, as well as three volunteers are impacted by the Launceston closure.
"The closure of trading operations in no way reflects the performance of our valued staff and volunteers over more than 50 years," Mr McKimmie said.
The Centreway Arcade store has been operating for more than 20 years. It will close on June 21.
"We sincerely thank (staff) - as well as our customers - for their dedication and significant contributions," Mr McKimmie said.