caa a22 4500 477137385 CHVBK 20180405111722.0 cr unu---uuuuu 170330e19971201xx s 000 0 eng 10.1023/A:1008840603393 doi (NATIONALLICENCE)springer-10.1023/A:1008840603393 Gln54Leu of the conserved polar residue inthe interfacial coiled coil position (d) results in significant stabilization of the original structure of the COMP pentamerization domain [Elektronische Daten] [Alexey Terskikh, Sergey Potekhin, Tatiana Melnik, Andrey Kajava] The assembly domain of cartilage oligomeric matrixprotein (COMP) forms an α-helical coiled coilhomopentamer with a conserved polar glutamine in theinterior (d) position. We substituted Gln54 forapolar Leu in the recombinant fragment of the rat COMPdomain. Biochemical studies and circular dichroism(CD) spectroscopy showed that the mutant, similarly tothe wild-type (w.t.) peptide, forms spontaneously anα-helical pentamer. Thermal transitions of thew.t. and mutant pentamers were analyzed by CDspectroscopy and differential scanning calorimetry.The Gln54Leu mutation increased the thermal stabilityof the pentamer with reduced disulfide bonds from73 °C to 104 °C. The denaturation of thedisulfide bonded w.t. pentamer was observed at108 °C while the mutant pentamer cannot bedenatured up to 120 °C (the apparatus limit).Thus, by Gln54Leu mutation we found a way tosignificantly stabilize the coiled coil pentamer,making this peptide even more attractive as an oligomerization tool for various biotechnological applications. Kluwer Academic Publishers, 1997 Cartilage oligomeric matrix protein nationallicence Coiled coils nationallicence Mutation nationallicence Pentamer nationallicence Thrombospondin nationallicence Terskikh Alexey Institute of Biochemistry, University of Lausanne, Ch. des Boveresses 155, CH-1066, Epalinges, Switzerland aut Potekhin Sergey Institute of Protein Research, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia aut Melnik Tatiana Institute of Protein Research, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia aut Kajava Andrey Swiss Institute for Experimental Cancer Research, Ch. des Boveresses 155, CH-1066, Epalinges, Switzerland aut Letters in Peptide Science Kluwer Academic Publishers 4/4-6(1997-12-01), 297-304 0929-5666 4:4-6<297 1997 4 10989 https://doi.org/10.1023/A:1008840603393 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1008840603393 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Terskikh Alexey Institute of Biochemistry, University of Lausanne, Ch. des Boveresses 155, CH-1066, Epalinges, Switzerland aut NATIONALLICENCE 700 1- Potekhin Sergey Institute of Protein Research, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia aut NATIONALLICENCE 700 1- Melnik Tatiana Institute of Protein Research, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia aut NATIONALLICENCE 700 1- Kajava Andrey Swiss Institute for Experimental Cancer Research, Ch. des Boveresses 155, CH-1066, Epalinges, Switzerland aut NATIONALLICENCE 773 0- Letters in Peptide Science Kluwer Academic Publishers 4/4-6(1997-12-01), 297-304 0929-5666 4:4-6<297 1997 4 10989 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer