Amino acids in segment IVS6 and beta-subunit interaction support distinct conformational changes during Ca(v)2.1 inactivation

Ca(v)2.1 mediates voltage-gated Ca2+ entry into neurons and the release of neurotransmitters at synapses of the central nervous system. An inactivation process that is modulated by the auxiliary beta-subunits regulates Ca2+ entry through Ca(v)2.1. However, the molecular mechanism of this alpha1-beta-subunit interaction remains unknown. Herein we report the identification of new determinants within segment IVS6 of the alpha(1)2.1-subunit that markedly influence channel inactivation. Systematic substitution of residues within IVS6 with amino acids of different size, charge, and polarity resulted in mutant channels with rates of fast inactivation (k(inact)) ranging from a 1.5-fold slowing in V1818I (k(inact) = 0.98 +/- 0.09 s(-1) compared with wild type alpha(1)2.1/alpha2-delta/beta1a k(inact) = 1.35 +/- 0.25 s(-1) to a 75-fold acceleration in mutant M1811Q (k(inact) = 102 +/- 3 s(-1). Coexpression of mutant alpha(1)2.1-subunits with beta(2a) resulted in two different phenotypes of current inactivation: 1) a pronounced reduction in the rate of channel inactivation or 2) an attenuation of a slow component in I(Ba) inactivation. Simulations revealed that these two distinct inactivation phenotypes arise from a beta2a-subunit-induced destabilization of the fast-inactivated state. The IVS6- and beta2a-subunit-mediated effects on Ca(v)2.1 inactivation are likely to occur via independent mechanisms.     

Happy orang-utans live longer lives

Nonhuman primate ageing resembles its human counterpart. Moreover, ratings of subjective well-being traits in chimpanzees, orang-utans and rhesus macaques are similar to those of humans: they are intercorrelated, heritable, and phenotypically and genetically related to personality. We examined whether, as in humans, orang-utan subjective well-being was related to longer life. The sample included 184 zoo-housed orang-utans followed up for approximately 7 years. Age, sex, species and number of transfers were available for all subjects and 172 subjects were rated on at least one item of a subjective well-being scale. Of the 31 orang-utans that died, 25 died a mean of 3.4 years after being rated. Even in a model that included, and therefore, statistically adjusted for, sex, age, species and transfers, orang-utans rated as being "happier" lived longer. The risk differential between orang-utans that were one standard deviation above and one standard deviation below baseline in subjective well-being was comparable with approximately 11 years in age. This finding suggests that impressions of the subjective well-being of captive great apes are valid indicators of their welfare and longevity.     

Integrin alpha v beta 3 binds a unique non-RGD site near the C-terminus of human tropoelastin

Tropoelastin is the soluble precursor of the essential resilient connective tissue protein elastin. We examined the binding of integrin alpha(v)beta(3) to tropoelastin. In quantitative colorimetric solid-phase assays, purified alpha(v)beta(3) demonstrated saturable, divalent cation-dependent, single-site binding behavior on tropoelastin with a dissociation constant of 3.8 +/- 0.9 nM in the presence of 1 mM Mn(2+) which increased to 23 +/- 5 nM in the presence of 1 mM Ca(2+). Association with alpha(v)beta(3) was localized to the C-terminal 16 residues of tropoelastin, encompassing the region encoded by exon 36. This region comprises a unique disulfide loop in tropoelastin that is not essential for the interaction. This is the first identification of a specific, single binding site on tropoelastin and the first observation of direct binding of an integrin to a tropoelastin domain.