Thrombospondin-1 (TSP-1) is a protein involved in angiogenesis and tumor metastasis. In a previous study, a tridecapeptide sequence of TSP-1B [KRFKQDGGWSHWG] was synthesized and its biological activity was determined as well as the activity of three related sequences TSPB-(E), TSPB-(S), and TSPB-(Abu)(6). These peptides were tested for activity on the cell growth of three human carcinoma cells lines and only TSPB-(Abu)(6) increased proliferation of MCF7 and HT-29. The main aim of this study was to perform physicochemical measurements, in a comparative way, to determine if the differences in activity could be related to physicochemical properties. Peptides were characterised by HPLC capacity factors, UV, fluorescence, and CD spectra (either in buffer solution or in the presence of lipid vesicles), surface activity, and aggregation. Moreover, the interaction of these peptides with phospholipids was determined through their penetration in monolayers of DPPC, PG, or PS as well as their miscibility in mixed monolayers. Besides, using liposomes as model membranes, the affinity of these peptides for phosphatidylcholine was measured with vesicles labeled with fluorescent markers (TMA-DPH, laurdan, pyrene). Results show that these molecules are highly hydrophilic and their surface activity is low. Mixed monolayers indicate that there is almost no miscibility. Besides, its presence does not modify noticeably the microviscosity of bilayers. Moreover, UV and fluorescence spectra of peptides were not affected by the presence of lipids in the media but CD spectra recorded in TFE/water (1/1) resulted in small changes for TSPB, TSPB-(E), and TSPB-(S) peptides. On the contrary CD spectra of TSPB-(Abu)(6) derivatives were clearly much more sensitive to the polarity of the environment. According to these data the biological activity of peptide with a cyclic aspartimide moiety at position 6 could be related to a specific conformational change in the peptide chain promoted by a hydrophobic membrane-like environment. (c) 2011 Elsevier Inc. All rights reserved.