The store in Barker’s Pool, near City Hall, which is also home to the Early Learning Centre, is set to close its doors for good on Wednesday, January 25.
The news was broken to shocked staff at the store, where 20 people work, on New Year’s Eve.
Branch manager Yvette Kendall blamed the store’s downturn on major disruption caused by widescale development in the city centre.
She said traffic and parking woes had deterred shoppers from visiting the area over the last six months.
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“The redevelopment has caused a lot of hassle, because customers have found it hard to get into town and park their cars,” she said.
“We know we have to go through this to get a redeveloped town centre but it has affected sales big time.
“Our customers have been shocked by the news. A lot of them first came here as young people and have come back as parents and now grandparents.”
Ms Kendall said famous customers had included Olympic great Jessica Ennis-Hill, who visited when she was pregnant.
The closest Mothercare branches will now be at Meadowhall and Crystal Peaks shopping centres.
A spokeswoman for Mothercare said the closure was part of the company’s ‘transformation plan’.
“We are working very closely with our team and every effort will be made to relocate colleagues at nearby stores where possible,” she said.
“Whilst we regret the closure of the store, it is part of our larger transformation plan for our business to close unprofitable stores and invest in a store estate that provides our customers with a consistently enjoyable shopping experience, combining great product and service with the flexibility and convenience to shop both in store and online.”