caa a22 4500 397558554 CHVBK 20180308164810.0 cr unu---uuuuu 161202e199611 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021502 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021502 Gelation of the Lens Capsule Type IV Collagen Solution at a Neutral pH [Elektronische Daten] [Koichi Nakazata, Masatoshi Muraoka, Eijiro Adachi, Toshihiko Hayashi] It is known that the type IV collagen extracted from EHS tumor assembles under a physiological condition, but not in a gel form. The EHS type IV collagen requires the other basement membrane components, laminin1, heparansulfate proteoglycan, and/or nidogen for gelation. On the other hand, Muraoka et al. reported that the bovine lens capsule type IV collagen alone gelated under a unique and unexpected condition of 2 M guanidine-HCl and 50 mM dithiothreitol, a condition which is thought to be dissociative for most biological macromolecules, including extracellular matrix [Muraoka, M. et al. (1996) J. Biochem. 119, 167-172]. The present report shows that the bovine lens capsule type IV collagen formed a gel under physiological conditions of pH and ionic environment, though the apparent rigidity of the gel was weaker than that of the gel formed in 2 M guanidine-HCl and dithiothreitol. The rigidity depended greatly on the incubation temperature and NaCl concentration of the type IV collagen solution, as observed in terms of the contractility of gel volume under centrifugal force. The gel formed in 150 mM NaCl and 20 mM phosphate, pH 7.3, at 28°C contracted to 20% of the original volume on centrifugation of 1,800× g for 10 min, while the gel formed at 4°C, where type I collagen did not gelate at all, retained 90% of the original volume at the same centrifugal force. NaCl concentration was another important factor influencing the mechanical properties of type IV collagen gel. The gel formed at 150 mM showed maximal rigidity in the range of 0 to 300 mM in terms of the contractility on centrifugation. An image of a Pt/C replica of the gelated type IV collagen reconstituted at 4 or 28°C in 20 mM phosphate, pH 7.3, containing 150 mM NaCl showed fine meshworks consisting of rather homogeneous pore sizes, resembling the skeletal structure of basal lamina. Since the condition where the type IV collagen alone formed gels was physiological in terms of ionic strength and pH, the aggregate structure and gel properties might reflect the in vivo type IV collagen supramolecular structure and the property. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence basement membrane nationallicence gel nationallicence lens capsule nationallicence self-assembly nationallicence type IV collagen nationallicence Nakazata Koichi Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo 3-8-1 Komaba, Meguro-ku, Tokyo 153 aut Muraoka Masatoshi The Tokyo Metropolitan Institute of Medical Science aut Adachi Eijiro Department of Cell Biology, Kitasato University School of Medicine aut Hayashi Toshihiko Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo 3-8-1 Komaba, Meguro-ku, Tokyo 153 aut The Journal of Biochemistry Oxford University Press 120/5(1996-11), 889-894 0021-924X 120:5<889 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021502 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021502 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nakazata Koichi Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo 3-8-1 Komaba, Meguro-ku, Tokyo 153 aut NATIONALLICENCE 700 1- Muraoka Masatoshi The Tokyo Metropolitan Institute of Medical Science aut NATIONALLICENCE 700 1- Adachi Eijiro Department of Cell Biology, Kitasato University School of Medicine aut NATIONALLICENCE 700 1- Hayashi Toshihiko Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo 3-8-1 Komaba, Meguro-ku, Tokyo 153 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/5(1996-11), 889-894 0021-924X 120:5<889 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