The objective of this study was to evaluate the effects of dietary fat source with modified distillers grains plus solubles (MDGS) on beef tenderness through examination of sarcoplasmic reticulum (SR) membrane fatty acid composition, changes in sarcoplasmic calcium concentration, Warner-Bratzler shear force (WBSF), sarcomere length, and proteolysis early postmortem. Steers (n = 256) were fed for 134 d on either corn, or a diet containing 40% full-fat MDGS, 40% de-oiled MDGS, or 38% de-oiled MDGS plus 2% corn oil. Twenty-four USDA Choice carcasses (three carcasses per pen; eight pens per treatment) were selected within each dietary treatment, and strip loins were collected and aged for 2, 9, 16, or 23 d postmortem and subject to retail display (RD) for an additional 7 d. Feeding MDGS increased (P < 0.05) concentrations of linoleic acid (18:2) and tended to increase (P = 0.06) total polyunsaturated fatty acids (PUFA) in the SR membrane. Steaks from cattle fed MDGS had greater sarcoplasmic calcium concentration than steaks from cattle fed corn at day 2 postmortem (P = 0.05). Steaks from steers fed de-oiled MDGS and de-oiled MDGS plus corn oil tended to have lower WBSF values (P = 0.08) than steaks from cattle fed corn at 2 d of aging with 0 d of RD. There were no differences among dietary treatments for sarcomere length (P = 0.92) and troponin-T degradation at 2 d postmortem (P = 0.60). Results suggest that feeding MDGS may increase early postmortem release of calcium into the sarcoplasm due to increased 18:2 and PUFA concentration in the SR membrane, which could result in increased beef tenderness early postmortem.