The association of paraoxonase-1 (PON1) with high-density lipoproteins (HDL) is a prerequisite for maintaining normal serum activity of the enzyme. The lipoprotein furnishes an amphipathic environment to shield the hydrophobic, N-terminal region of the enzyme, and such an environment may also be necessary for interaction of PON1 with its substrates. HDL provides the optimal physiological acceptor complex, in terms of both stimulating PON1 secretion and stabilizing the secreted peptide. Lipid and peptide components of HDL contribute to these effects, such that modulating HDL composition influences PON1 activity and function. In this context, understanding how PON1 associates with HDL, what governs the association, and the mechanism by which the PON1-HDL complex exerts its antioxidant function is of particular physiological relevance. Moreover, HDL is subject to substantial compositional variations under both normal and pathological metabolic conditions. It has implications for the influence of the enzyme on cardiovascular risk, as normal enzyme activity may not correlate with optimal functional (antioxidant) efficiency. We review evidence that HDL lipid and protein components interact to promote PON1 secretion and maintain serum enzyme activity. Emerging data on how the enzyme associates with HDL are discussed, and the consequences for PON1 function of modifications to HDL are outlined. Finally, we highlight questions concerning the HDL-PON1 association that remain unanswered but are of particular importance in defining PON1 efficiency.