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Crossbred steers (n = 136) were used to assess breed differences in growth performance, carcass characteristics, fatty acid composition (total lipids and phospholipids), and palatability attributes of longissimus muscle. A multiple regression model was applied to crossbreeding data to estimate genetic differences between Simmental and Red Angus at the same level of backfat finish (10 mm). Simmental spent 71 more (P < 0.001) days on feed to acquire the same degree of backfat thickness as Red Angus, had heavier (P < 0.001) slaughter weights, larger (P = 0.002) longissimus muscle area, and increased (P = 0.023) lean yield. Average daily gain did not differ (P = 0.297) between breeds. Simmental were less (P = 0.012) efficient in converting feed to gain than Red Angus. Generally, there were few breed differences in palatability attributes for longissimus and semitendinosus muscles, with the exception of increased (P < 0.05) beef flavor scores for Simmental beef vs Red Angus beef across both muscles. For total lipids, concentrations of myristoleic acid (14:1), palmitoleic acid (16:1), and vaccenic acid (18:1n-7), along with n-6 to n-3 fatty acid (n-6:n-3) ratio, were greater (P < 0.05) in Simmental than Red Angus. In contrast, concentrations of margaric acid (17:0), eicosapentaenoic acid (20:5n-3), and total n-3 polyunsaturated fatty acids (n-3 PUFA) were greater (P < 0.05) in Red Angus than Simmental. For phospholipids, Simmental had lower (P < 0.05) amounts of 20:5n-3, docosahexaenoic acid (22:6n-3), and n-3 PUFA, with a greater (P = 0.017) n-6:n-3 ratio. Activity of delta9-desaturase enzyme in the conversion of palmitic acid (16:0) to 16:1 was greater (P = 0.001) in total lipids from Simmental as compared with Red Angus. A genetic basis for fatty acid differences is suggested, although the biological and practical significance needs to be demonstrated.