caa a22 4500 477137989 CHVBK 20180405111724.0 cr unu---uuuuu 170330e19971201xx s 000 0 eng 10.1007/BF02442893 doi (NATIONALLICENCE)springer-10.1007/BF02442893 Mutation Gln54Leu of the conserved polar residue in the 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] Summary: The assembly domain of cartilage oligomeric matrix protein (COMP) forms an α-helical coiled coil homopentamer with a conserved polar glutamine in the interior (d) position. We substituted Gln54 for apolar Leu in the recombinant fragment of the rat COMP domain. Biochemical studies and circular dichroism (CD) spectroscopy showed that the mutant, similarly to the wild-type (w.t.) peptide, forms spontaneously an α-helical pentamer. Thermal transitions of the w.t. and mutant pentamers were analyzed by CD spectroscopy and differential scanning calorimetry. The Gln54Leu mutation increased the thermal stability of the pentamer with reduced disulfide bonds from 73°C to 104°C. The denaturation of the disulfide bonded w.t. pentamer was observed at 108°C while the mutant pentamer cannot be denatured up to 120°C (the apparatus limit). Thus, by Gln54Leu mutation we found a way to significantly 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/10.1007/BF02442893 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02442893 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