The US Consulate in Chengdu was closed on Monday after China asked it to be shut down last week as a retaliatory move against Washington's decision to close the Asian giant's consulate in Houston.
A US State Department statement said that the Consulate suspended operations at 10 am on Monday.
China had given the US time till the 27th of July to close the Consulate.
Even as the US closed the Consulate, it expressed disappointment at China's decision.
The State Department said the US would try to continue its outreach to the region through its other missions in China.
The Chengdu Consulate "has stood at the center of our relations with the people in Western China, including Tibet, for 35 years", it added.
The US-China tensions have escalated since President Donald Trump's administration renegotiated terms of trade with the regime of his counterpart, Xi Jinping.
The two countries entered into a cold war-like situation after China withheld information about the coronavirus pandemic, resulting in a global crisis.