Adipocyte-derived mediators contribute to chronic, diabetes-associated inflammation. We recently demonstrated, that heat shock protein 60 (Hsp60) is an effective inductor of inflammatory adipocyte activities. In the present study, we characterized the initial Hsp60 binding to adipocyte receptor structures. Analyses with preadipocytes and adipocytes from the murine 3T3-L1 line and with primary cultures from the New Zealand obese mouse, a model of human obesity, revealed comparable specific, dose-dependent and saturable Hsp60 binding, confirming the characteristics of a ligand-receptor interaction. Furthermore, we identified the N-terminal regions aa1-50 and aa91-110 of the Hsp60 molecule as relevant epitopes involved in binding to receptor structures on these cells. Our results demonstrate differentiation-independent conserved Hsp60 reactivity in permanent and primary adipocytes, strongly indicating that Hsp60 is an important regulator of inflammatory adipocyte activities. (C) 2010 Elsevier Inc. All rights reserved.