The fashion chain will close in April and follow Debenhams out of the city.
Topshop has been among the most popular shops on Dudley Street over the years but has now been revealed as one of the branches to be axed by Sir Philip Green's struggling retailer.
The billionaire tycoon's Arcadia Group agreed to go ahead with a Company Voluntary Arrangement (CVA) in the summer resulting in the closure of some of its Topshop and Dorothy Perkins stores.
Topshop reported a £500m loss last year amid falling sales.
Documents discussing the future of the Dudley Street unit revealed it will close in April.
They said: "The unit is occupied by Topshop /Topman however they are currently under a Company Voluntary Arrangement (CVA) for closure in April 2020.
"The property has been marketed since July 2019 following the CVA approval in June."
However, it appears the unit will not be left empty for long as there are already plans to turn it into a bistro and bar.
National chain Loungers has lodged plans to launch once Topshop has moved out.
The loss of the retailer delivers a fresh blow to Wolverhampton city centre which had been enjoying something of a revival following the move of Wilko's to the Mander Centre and opening of Matalan.
Debenhams, which opened as the anchor store at the newly-refurbished Mander Centre two years ago, will close in the new year.
Loungers said its plan for the bistro and bar would "enhance the provision of eating and drinking establishments" in the city centre while providing jobs and "significant investment".
Wendy Thompson, Conservative leader in the city, said the loss of Topshop would be a blow.
She said: "It's considerably disappointing. It's very important to try and keep a reasonable amount of choice of retail shops, it adds to the vibrancy of the city centre.
"It is good that is going to be used because it would be very detrimental if we were to have another empty building."