James led the Gators in kickoff and punt return yardage for four consecutive seasons, and finished his college career with 4,770 total yards gained, including 112 kickoff returns for 2,718 yards and a touchdown, 117 punt returns for 1,371 yards and four touchdowns, 379 receiving yards and 302 rushing yards. His career kickoff return yardage and combined kick return yardage remain SEC records. He was a first-team All-American in 2007, a consensus first-team All-American in 2008, a first-team All-SEC selection in 2008, the SEC Special Teams Player of the Year in 2008, and a senior team captain in 2009. During his time as a Gator, the team won two SEC Championship Games (2006, 2008) and two BCS National Championship Games (2007, 2009).
The Indianapolis Colts signed James as an undrafted free agent on April 30, 2010. On November 10, James was activated from the practice squad to the main roster. On November 28, 2010, James posted 150 kick return yards and 29 punt return yards. He was released the following week from the Colts active roster.
In 2011, the Edmonton Eskimos signed James as a free agent, but he was later released during the regular season, bringing his playing career to an end.
In 2012, James was hired to coach the football team at St. Joseph Academy in St. Augustine. He was fired after one season in which the team went 2-8.