caa a22 4500 397558597 CHVBK 20180308164810.0 cr unu---uuuuu 161202e199611 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021509 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021509 Separation of the Subtypes of Type V Collagen Molecules, [αl(V)]2α2(V) and α1(V)α2(V)α3(V), by Chain Composition-Dependent Affinity for Heparin: Single α1(V) Chain Shows Intermediate Heparin Affinity between Those of the Type V Collagen Subtypes Composed of [α1(V)]2α2(V) and of α1(V)α2(V)α3(V) [Elektronische Daten] [Kazunori Mizuno, Toshihiko Hayashi] The heparin affinities of heat-treated type V collagen α-chains and the triple-helical molecules were evaluated in terms of the NaCl concentration required for prevention of binding to a heparin-Sepharose column. After heat treatment, α1(V) chain required approximately two-fold higher NaCl concentration to pass through the column than the other two chains, α2(V) and α3(V). Thus, the heparin affinity of α1(V) may be approximately two-fold higher than those of the other α(V)-chains. The type V collagen molecules in triple-helical conformation were separated into two fractions at 170 mM NaCl in 20 mM phosphate buffer, pH 7.2, containing 2 M urea; bound and non-bound. The ratio of the three a-chains, αl(V):α2(V):α3(V) was 2: 1 : 0 and 1: 1: 1 in the bound and flow-through fractions, respectively, on analysis by SDS-PAGE. The differential affinity of the two fractions could be accounted for by the number of α1(V) chains in the triple-helical molecule, if these fractions contained triple-helical subtypes with the chain compositions of [α1(V)]2α2(V) and α1(V)α2(V)α3(V), respectively. From the comparison of the NaCl concentration required for prevention of the binding, [α1(V)]2α2(V) had about two-fold higher affinity than α1(V)α2(V)α3(V), and the separated α1(V) chain showed an intermediate affinity. A possible explanation for difference in heparin affinity among the subtypes of molecules and the separated α-chains is that the heparin affinity of type V collagen molecule is governed by the number of α1(V) chains contained in the molecule and that steric restraint in a triple-helical conformation weakens the binding of αl(V) chain to heparin. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence affinity chromatography nationallicence heparin binding nationallicence heparin column nationallicence triple-helical structure nationallicence type V collagen nationallicence Mizuno Kazunori Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Kornaba 3-8-1, Meguro-ku, Tokyo 153 aut Hayashi Toshihiko Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Kornaba 3-8-1, Meguro-ku, Tokyo 153 aut The Journal of Biochemistry Oxford University Press 120/5(1996-11), 934-939 0021-924X 120:5<934 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021509 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021509 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mizuno Kazunori Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Kornaba 3-8-1, Meguro-ku, Tokyo 153 aut NATIONALLICENCE 700 1- Hayashi Toshihiko Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Kornaba 3-8-1, Meguro-ku, Tokyo 153 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/5(1996-11), 934-939 0021-924X 120:5<934 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford