Human Calmodulin-Like Skin Protein (CLSP) is a 15.92 kDa protein expressed during the differentiation of keratinocytes to corneocytes. The structure and dynamics of this protein have been characterized by NMR spectroscopy. CLSP is affected by high mobility both in the fast (ps-ns) and intermediate (µs) timescale range, with the N-terminal and C-terminal domains being differently affected. The isolated N-terminal and C-terminal domains were also expressed and analyzed. The structure of the isolated C-terminal domain is presented, which is very similar to that in the entire protein. The N-terminal domain is characterized by four stable helices which experience large fluctuations. This is shown to be due to mutations in the hydrophobic core. Fast mobility is experienced in the two loops. Almost no long range NOEs are observed. The overall N-terminal domain behavior is similar both in the full length protein and in the isolated domain. By exploiting the capability of Tb3+ bound to CLSP to induce partial orientation of the molecule in a magnetic field, restricted motion of one domain with respect to the other was proved. By using NMR, ITC and ESI-MS the calcium and magnesium binding properties were investigated. Finally CLSP is framed into the evolutionary scheme of the Calmodulin-like family
Babini E., Bertini I., Capozzi F., Chirivino E., Luchinat C. (2006). A structural and dynamic characterization of the EF-hand protein CLSP. STRUCTURE, 14, 1029-1038 [10.1016/j.str.2006.04.004].
A structural and dynamic characterization of the EF-hand protein CLSP
BABINI, ELENA;CAPOZZI, FRANCESCO;
2006
Abstract
Human Calmodulin-Like Skin Protein (CLSP) is a 15.92 kDa protein expressed during the differentiation of keratinocytes to corneocytes. The structure and dynamics of this protein have been characterized by NMR spectroscopy. CLSP is affected by high mobility both in the fast (ps-ns) and intermediate (µs) timescale range, with the N-terminal and C-terminal domains being differently affected. The isolated N-terminal and C-terminal domains were also expressed and analyzed. The structure of the isolated C-terminal domain is presented, which is very similar to that in the entire protein. The N-terminal domain is characterized by four stable helices which experience large fluctuations. This is shown to be due to mutations in the hydrophobic core. Fast mobility is experienced in the two loops. Almost no long range NOEs are observed. The overall N-terminal domain behavior is similar both in the full length protein and in the isolated domain. By exploiting the capability of Tb3+ bound to CLSP to induce partial orientation of the molecule in a magnetic field, restricted motion of one domain with respect to the other was proved. By using NMR, ITC and ESI-MS the calcium and magnesium binding properties were investigated. Finally CLSP is framed into the evolutionary scheme of the Calmodulin-like familyI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
