The present study was conducted to test the association between protein intake and blood pressure, glucose levels, and blood cholesterol in a large sample of Italian older adults. Longevity Check-up 7+ (Lookup 7+) is an ongoing project that started in June 2015. The project is conducted in unconventional settings (e.g., exhibitions, malls, health promotion campaigns) across Italy with the aim of fostering adoption of healthy lifestyles in the general population. For the present study, participants were eligible if they were 65+ years and provided written informed consent. Systolic (SBP) and diastolic blood pressure (DBP), and blood glucose and cholesterol levels were assessed. Protein intake was estimated using a 12-item food frequency questionnaire. Three-thousand four-hundred and four older adults were included in the study. The results of the linear regression showed an inverse association between protein intake (as a continuous variable) and DBP, and a positive correlation with blood cholesterol levels. The findings of the present study indicate that a high intake of protein was negatively associated with DBP and positively associated with total blood cholesterol levels in a large cohort of Italian older adults, after adjustment for numerous covariates.