Objective: Latinos disproportionately experience overweight/obesity (OWOB) and insufficient physical activity (PA), which are risk factors for numerous health conditions. Whereas numerous studies investigate acculturation as a determinant of OWOB and PA, few have examined acculturation multidimensionally, and none has examined its interaction with gender. Methods: Participants were 140 Latino adults. Primary outcomes were status as OWOB and endorsement of insufficient PA. Acculturation was measured with the Multidimensional Acculturation Scale II. Logistic regression analyses were used to examine the moderating effect of gender on the relationship between each acculturation scale and outcome, controlling for education, nativity, and smoking status. Results: Greater Spanish proficiency was significantly associated with lower odds of insufficient PA. Greater American Cultural Identity was significantly associated with higher odds of OWOB. Women had significantly higher odds of endorsing insufficient PA compared to men. Gender did not moderate the relationship between acculturation and either OWOB or PA. Conclusions: Acculturation is similarly associated with OWOB and insufficient PA for Latino men and women. Cultural identity may need to be considered to target OWOB interventions. Acculturation may be less important, and gender more important, to consider for appropriate targeting of PA interventions.