In this paper, a new in situ method for determining the structural rolling resistance (SRR), defined as the dissipated energy caused by deformation of the pavement when subjected to a moving load, is presented. The method is based on the relation between SRR and the slope of the deflection basin under a moving load. Using the Traffic Speed Deflectometer, the deflection slope is measured at several positions behind and in front of the right rear-end tire pair of a full-size truck trailer while driving under realistic conditions. The deflection slope directly under the tire is estimated from a linear interpolation between the two nearest sensors. A set of data from a test road segment located in Denmark is analyzed and the SRR coefficients are found to be in the range 0.005% to 0.05%. The deflection slope measurements have a high reproducibility (repeated measurements agree within standard deviations of 4% to 10%) with high spatial resolution, and the method for calculating SRR from these measurements has the clear advantage that it requires no knowledge or model of the pavement structure or viscoelastic properties. Numerical simulations of pavement response show that the proposed interpolation method tends to underestimate the actual SRR, and better estimates can be obtained by other interpolation schemes.