Trufant was also a standout long jumper and triple jumper on the Rams’ track team, and took second place in the Class 4A state track meet in the triple jump (46 feet, 5.75 inches) and 11th in the long jump (21 feet, 2.25 inches) as a junior in 2008. He also played basketball at Wilson, earning second-team all-league as a junior.

The Atlanta Falcons selected Trufant in the first round (22nd overall) of the 2013 NFL Draft. He was the third cornerback selected in 2013, behind Alabama’s Dee Milliner (9th overall) and Houston’s D. J. Hayden (12th overall), and was also the first of two cornerbacks the Falcons selected, along with second round pick Robert Alford (60th overall).

On July 24, 2013, the Atlanta Falcons signed Trufant to a four-year, $8.16 million contract that includes $6.94 million guaranteed and a signing bonus of $4.31 million.

He made his professional regular season debut and first career start in the Atlanta Falcons’ season-opener at the New Orleans Saints and made five combined tackles and a pass deflection in a 23–17 loss. In Week 7, Trufant recorded a season-high seven combined tackles and broke up a pass during a 31–23 victory against the Tampa Bay Buccaneers. On November 3, 2013, Trufant recorded five combined tackles, a pass deflection, and his first career interception off a pass by quarterback Cam Newton in the Falcons’ 34–10 loss at the Carolina Panthers. On December 15, 2013, Trufant tied his season-high with seven combined tackles, two pass deflections, and intercepted a pass by quarterback Kirk Cousins in their 27–26 victory against the Washington Redskins. He finished his rookie season in 2013 with a career-high 70 combined tackles (55 solo), a career-high 17 pass deflections, two interceptions, one forced fumble, and two fumble recoveries in 16 games and 16 starts. He exceeded expectations, being ranked eighth out of all cornerbacks in the league by Pro Football Focus and allowed a 53.% completion percentage and a quarterback rating of 75.2 to opposing quarterbacks in pass coverage.

Trufant became the Falcons’ No. 1 cornerback after they released Asante Samuel. He was officially named the starting cornerback, alongside Robert Alford, to begin the regular season. On November 9, 2014, Trufant recorded a season-high seven combined tackles and a pass deflection during their 27–17 victory at the Tampa Bay Buccaneers. The following week, he made a season-high two pass deflections and intercepted a pass by Cam Newton in a 19–17 victory at the Carolina Panthers. On December 29, 2014, the Atlanta Falcons fired head coach Mike Smith after the Falcons did not qualify for the playoffs after finishing with a 4–12 record. Trufant finished the 2014 season with 61 combined tackles (53 solo), 16 pass deflections, and three interceptions in 16 games and 16 starts.

He started the Atlanta Falcons’ season-opener against the Tampa Bay Buccaneers and collected a season-high five combined tackles, broke up a pass, and intercepted a pass by Jameis Winston during a 31–24 loss. On October 16, 2016, Trufant made five combined tackles and a sack on Russell Wilson in the Falcons’ 26-24 loss at the Seattle Seahawks. In Week 9, Trufant made four solo tackles before leaving the Falcons’ 43–28 victory against the Tampa Bay Buccaneers in the first half after suffering a shoulder injury. On November 29, 2016, the Atlanta Falcons placed Trufant on injured reserve after they discovered he would require shoulder surgery. Pro Football Focus gave Trufant an overall grade of 78.9, ranking 30th among all qualifying cornerbacks in 2016. He finished the 2016 season with 31 combined tackles (25 solo), four pass deflections, two sacks, and an interception in nine games and nine starts.

The Atlanta Falcons went to Super Bowl LI after finishing atop the NFC South with an 11–5 record and defeating the Seattle Seahawks in the NFC Divisional Round and the Green Bay Packers in the NFC Championship. On February 5, 2017, the Atlanta Falcons lost 34–28 in overtime to the New England Patriots in Super Bowl LI after leading 28–9 going into the fourth quarter.

On April 8, 2017, the Atlanta Falcons signed Trufant to a five-year, $68.5 million contract that includes $41.52 million and $15 million signing bonus.

New defensive coordinator Marquand Manuel who replaced Rick Smith after the Falcons’ collapse in Super Bowl LI. In Week 2, Trufant recorded two solo tackles, broke up a pass, an interception, and recovered a fumble off a strip sack by Vic Beasley and returned it 15-yards for a touchdown during a 34–23 victory against the Green Bay Packers on NBC Sunday Night Football. His performance earned him NFC Defensive Player of the Week. On November 20, 2017, Trufant made five solo tackles, a season-high three pass deflections, and intercepted a pass by Russell Wilson during a 34–31 victory at the Seattle Seahawks. Trufant was inactive for the Falcons’ Week 13 loss to the Minnesota Vikings due to a concussion he received he previous week. He finished the 2017 season with 41 combined tackles (36 solo), 12 pass deflections, two interceptions, a sack, a fumble recovery, and a touchdown in 15 games and 15 starts. Pro Football Focus gave Trufant an overall grade of 84.9, which ranked 19th among all qualified cornerbacks in 2017.

In 2018, Trufant started all 16 games, recording 66 tackles, one sack, a team-high 12 passes defensed, and no interceptions.

On March 18, 2020, Trufant was released by the Falcons after seven seasons.

On March 25, 2020, Trufant signed a two-year, $21 million contract with the Detroit Lions.