Gene Expression of Neurotrophins and Their Receptors in Keloids.

The aim of this study was to assess gene expression of neurotrophins and their receptors in keloids. Skin samples of normal skin and keloids were obtained from patients in the control (n = 12) and keloid (n = 12) groups, respectively. Ribonucleic acid was extracted from the skin specimens, purified, evaluated by spectrophotometry, and used to synthesize complementary DNA. Real-time quantitative polymerase chain reaction analysis of 84 human neurotrophin genes and their receptors was performed. Twelve genes, including heat shock 27-kDa protein 1, gastrin-releasing peptide receptor, corticotropin-releasing hormone receptor 2, neuropeptide Y Y2 receptor, interleukin 6 signal transducer, nerve growth factor, metallothionein 3, B-cell chronic lymphocytic leukemia/lymphoma 2, cholecystokinin A receptor, persephin, galanin receptor 2, and fibroblast growth factor receptor 3, were down-regulated in keloid tissue compared with normal skin. The genes 27-kDa heat shock protein 1, gastrin-releasing peptide receptor, corticotropin-releasing hormone receptor 2, nerve growth factor, metallothionein 3, B-cell chronic lymphocytic leukemia/lymphoma 2, and persephin protein were considered priority genes associated with keloid formation.

Structural analysis of three peptides related to the transmambranic helix VI of AT1 receptor.

INTRODUCTION: Angiotensin II (AII) is the main active product of the renin angiotensin system. Better known effects of AII are via AT1 receptor (AT1R). Expression of AT1R mutants (L265D and L262D) in CHO cells increased cAMP formation when compared to CHO cells expressing the wild type (WT) AT1R. Morphological transformation of CHO cells transfected with mutants correlated with their increased cAMP formation. DNA synthesis was inhibited in these cells too, indicating that cAMP promotes inhibitory effects on transfected CHO cells growth and causes their morphological change from a tumorigenic phenotype to a non-tumorigenic one. OBJECTIVES: To assess the importance of leucine 262 and 265 in determining AT1R structure by means of a comparative structural analysis of two mutant peptides and of a wild-type fragment. METHODOLOGY: Three peptides had their conformation compared by circular dichroism (CD): L262D(259-272), L265D(259-272) (mutants) and WT(260-277). RESULTS: Secondary structures were: beta-turn for WT and L262D and random coil for L265D. CONCLUSIONS: Strong correlation was found in the results of biochemical, cellular and structural approaches used to compare WT AT1R to mutant types. Random coil structure of the L265D mutant may be a key point to explain those changes observed in biochemical (binding and signal transduction) and proliferation assays (Correa et al., 2005). beta-Turn formation is an important step during early protein folding and this secondary simple structure is present in L262D and WT, but not in L265D. Therefore, leucine 265 seems to play a crucial role in determining an entirely functional AT1R.