The agreement facilitates a potential deployment of US troops to the Czech Republic, a NATO member since 1999, said Defence Minister Jana Cernochova.
It also concerns the joint defence of NATO's eastern flank, logistical support to NATO allies and military training cooperation, she told reporters.
"The agreement will enable more intensive practical cooperation between the Czech army and the US armed forces," Cernochova added.
She said however that any presence of US troops on Czech territory would still have to be approved by the parliament.
Prime Minister Petr Fiala hailed the Defence Cooperation Agreement (DCA) as "undoubtedly a step that will increase the security of our country".
Prague and Washington are negotiating a date to sign the agreement.
The deal still requires approval from the Czech parliament, in which government parties have a majority, as well as President Petr Pavel, a former NATO general.
The Czech Republic has provided Ukraine with military aid worth $264 million since Russia's invasion began in February 2022, according to defence ministry data.
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