Agent Ricky Regufe was involved with the national team for 20 years and part of its delegation.
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He was also the key to Ronaldo‘s big-money move to Al-Nassr after the ex-Manchester United hero split from long-term representative Jorge Mendes.
It comes as Roberto Martinez prepares for his first two games as Portugal boss.
And Regufe’s announcement also follows speculation over the captaincy – long held by Ronaldo
Centre-back Pepe, 40, was listed as skipper at the recent FIFA Best Awards.
But Martinez has yet to make any decision on the matter public.
On stepping down from his Portugal position, Regufe posted: “It was a pride and a privilege to have served my country as a member of the National Team over two decades.
“I lived unforgettable moments and made friendships for life.
“I saw up close the National Team winning the two major trophies it has won so far and I watched Cristiano Ronaldo become the best player in the world.
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“I thank the players, coaches and staff for everything we’ve experienced together. A new stage begins but the desire to see our National Team win is always the same!”
Regufe added a picture of Portugal triumphing at Euro 2016, which was followed by winning the National League three years later.
Meanwhile, 38-year-old Ronaldo remains in Martinez’s plans despite bowing out of the World Cup in tears after only coming on as a sub in the last two games, including a shock quarter-final loss to Morocco.
Portugal manager Fernando Santos stepped down following that humiliation.