Effects of fibrillin-1 degradation on microfibril ultrastructure

Chiu Liang Kuo, Zenzo Isogai, Douglas R. Keene, Noriko Hazeki, Robert N. Ono, Gerhard Sengle, Hans Peter Bächinger, Lynn Sakai

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Current models of the elastic properties and structural organization of fibrillin-containing microfibrils are based primarily on microscopic analyses of microfibrils liberated from connective tissues after digestion with crude collagenase. Results presented here demonstrate that this digestion resulted in the cleavage of fibrillin-1 and loss of specific immunoreactive epitopes. The proline-rich region and regions near the second 8-cysteine domain in fibrillin-1 were easily cleaved by crude collagenase. Other sites that may also be cleaved during microfibril digestion and extraction were identified. In contrast to collagenase-digested microfibrils, guanidine-extracted microfibrils contained all fibrillin-1 epitopes recognized by available antibodies. The ultrastructure of guanidine-extracted microfibrils differed markedly from that of collagenase-digested microfibrils. Fibrillin-1 filaments splayed out, extending beyond the width of the periodic globular beads. Both guanidine-extracted and collagenase-digested microfibrils were subjected to extensive digestion by crude collagenase. Collagenase digestion of guanidine-extracted microfibrils removed the outer filaments, revealing a core structure. In contrast to microfibrils extracted from tissues, cell culture microfibrils could be digested into short units containing just a few beads. These data suggest that additional cross-links stabilize the long beaded microfibrils in tissues. Based on the microfibril morphologies observed after these experiments, on the crude collagenase cleavage sites identified in fibrillin-1, and on known antibody binding sites in fibrillin-1, a model is proposed in which fibrillin-1 molecules are staggered in microfibrils. This model further suggests that the N-terminal half of fibrillin-1 is asymmetrically exposed in the outer filaments, whereas the C-terminal half of fibrillin-1 is present in the interior of the microfibril.

Original languageEnglish (US)
Pages (from-to)4007-4020
Number of pages14
JournalJournal of Biological Chemistry
Volume282
Issue number6
DOIs
StatePublished - Jan 9 2007

Fingerprint

Microfibrils
Collagenases
Degradation
Guanidine
Digestion
Epitopes
Tissue
Antibody Binding Sites
Tissue culture
Fibrillin-1
Proline
Cysteine
Molecules
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kuo, C. L., Isogai, Z., Keene, D. R., Hazeki, N., Ono, R. N., Sengle, G., ... Sakai, L. (2007). Effects of fibrillin-1 degradation on microfibril ultrastructure. Journal of Biological Chemistry, 282(6), 4007-4020. https://doi.org/10.1074/jbc.M606370200

Effects of fibrillin-1 degradation on microfibril ultrastructure. / Kuo, Chiu Liang; Isogai, Zenzo; Keene, Douglas R.; Hazeki, Noriko; Ono, Robert N.; Sengle, Gerhard; Bächinger, Hans Peter; Sakai, Lynn.

In: Journal of Biological Chemistry, Vol. 282, No. 6, 09.01.2007, p. 4007-4020.

Research output: Contribution to journalArticle

Kuo, CL, Isogai, Z, Keene, DR, Hazeki, N, Ono, RN, Sengle, G, Bächinger, HP & Sakai, L 2007, 'Effects of fibrillin-1 degradation on microfibril ultrastructure', Journal of Biological Chemistry, vol. 282, no. 6, pp. 4007-4020. https://doi.org/10.1074/jbc.M606370200
Kuo CL, Isogai Z, Keene DR, Hazeki N, Ono RN, Sengle G et al. Effects of fibrillin-1 degradation on microfibril ultrastructure. Journal of Biological Chemistry. 2007 Jan 9;282(6):4007-4020. https://doi.org/10.1074/jbc.M606370200
Kuo, Chiu Liang ; Isogai, Zenzo ; Keene, Douglas R. ; Hazeki, Noriko ; Ono, Robert N. ; Sengle, Gerhard ; Bächinger, Hans Peter ; Sakai, Lynn. / Effects of fibrillin-1 degradation on microfibril ultrastructure. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 6. pp. 4007-4020.
@article{817bdff257bd4fb589c0990183da799d,
title = "Effects of fibrillin-1 degradation on microfibril ultrastructure",
abstract = "Current models of the elastic properties and structural organization of fibrillin-containing microfibrils are based primarily on microscopic analyses of microfibrils liberated from connective tissues after digestion with crude collagenase. Results presented here demonstrate that this digestion resulted in the cleavage of fibrillin-1 and loss of specific immunoreactive epitopes. The proline-rich region and regions near the second 8-cysteine domain in fibrillin-1 were easily cleaved by crude collagenase. Other sites that may also be cleaved during microfibril digestion and extraction were identified. In contrast to collagenase-digested microfibrils, guanidine-extracted microfibrils contained all fibrillin-1 epitopes recognized by available antibodies. The ultrastructure of guanidine-extracted microfibrils differed markedly from that of collagenase-digested microfibrils. Fibrillin-1 filaments splayed out, extending beyond the width of the periodic globular beads. Both guanidine-extracted and collagenase-digested microfibrils were subjected to extensive digestion by crude collagenase. Collagenase digestion of guanidine-extracted microfibrils removed the outer filaments, revealing a core structure. In contrast to microfibrils extracted from tissues, cell culture microfibrils could be digested into short units containing just a few beads. These data suggest that additional cross-links stabilize the long beaded microfibrils in tissues. Based on the microfibril morphologies observed after these experiments, on the crude collagenase cleavage sites identified in fibrillin-1, and on known antibody binding sites in fibrillin-1, a model is proposed in which fibrillin-1 molecules are staggered in microfibrils. This model further suggests that the N-terminal half of fibrillin-1 is asymmetrically exposed in the outer filaments, whereas the C-terminal half of fibrillin-1 is present in the interior of the microfibril.",
author = "Kuo, {Chiu Liang} and Zenzo Isogai and Keene, {Douglas R.} and Noriko Hazeki and Ono, {Robert N.} and Gerhard Sengle and B{\"a}chinger, {Hans Peter} and Lynn Sakai",
year = "2007",
month = "1",
day = "9",
doi = "10.1074/jbc.M606370200",
language = "English (US)",
volume = "282",
pages = "4007--4020",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "6",

}

TY - JOUR

T1 - Effects of fibrillin-1 degradation on microfibril ultrastructure

AU - Kuo, Chiu Liang

AU - Isogai, Zenzo

AU - Keene, Douglas R.

AU - Hazeki, Noriko

AU - Ono, Robert N.

AU - Sengle, Gerhard

AU - Bächinger, Hans Peter

AU - Sakai, Lynn

PY - 2007/1/9

Y1 - 2007/1/9

N2 - Current models of the elastic properties and structural organization of fibrillin-containing microfibrils are based primarily on microscopic analyses of microfibrils liberated from connective tissues after digestion with crude collagenase. Results presented here demonstrate that this digestion resulted in the cleavage of fibrillin-1 and loss of specific immunoreactive epitopes. The proline-rich region and regions near the second 8-cysteine domain in fibrillin-1 were easily cleaved by crude collagenase. Other sites that may also be cleaved during microfibril digestion and extraction were identified. In contrast to collagenase-digested microfibrils, guanidine-extracted microfibrils contained all fibrillin-1 epitopes recognized by available antibodies. The ultrastructure of guanidine-extracted microfibrils differed markedly from that of collagenase-digested microfibrils. Fibrillin-1 filaments splayed out, extending beyond the width of the periodic globular beads. Both guanidine-extracted and collagenase-digested microfibrils were subjected to extensive digestion by crude collagenase. Collagenase digestion of guanidine-extracted microfibrils removed the outer filaments, revealing a core structure. In contrast to microfibrils extracted from tissues, cell culture microfibrils could be digested into short units containing just a few beads. These data suggest that additional cross-links stabilize the long beaded microfibrils in tissues. Based on the microfibril morphologies observed after these experiments, on the crude collagenase cleavage sites identified in fibrillin-1, and on known antibody binding sites in fibrillin-1, a model is proposed in which fibrillin-1 molecules are staggered in microfibrils. This model further suggests that the N-terminal half of fibrillin-1 is asymmetrically exposed in the outer filaments, whereas the C-terminal half of fibrillin-1 is present in the interior of the microfibril.

AB - Current models of the elastic properties and structural organization of fibrillin-containing microfibrils are based primarily on microscopic analyses of microfibrils liberated from connective tissues after digestion with crude collagenase. Results presented here demonstrate that this digestion resulted in the cleavage of fibrillin-1 and loss of specific immunoreactive epitopes. The proline-rich region and regions near the second 8-cysteine domain in fibrillin-1 were easily cleaved by crude collagenase. Other sites that may also be cleaved during microfibril digestion and extraction were identified. In contrast to collagenase-digested microfibrils, guanidine-extracted microfibrils contained all fibrillin-1 epitopes recognized by available antibodies. The ultrastructure of guanidine-extracted microfibrils differed markedly from that of collagenase-digested microfibrils. Fibrillin-1 filaments splayed out, extending beyond the width of the periodic globular beads. Both guanidine-extracted and collagenase-digested microfibrils were subjected to extensive digestion by crude collagenase. Collagenase digestion of guanidine-extracted microfibrils removed the outer filaments, revealing a core structure. In contrast to microfibrils extracted from tissues, cell culture microfibrils could be digested into short units containing just a few beads. These data suggest that additional cross-links stabilize the long beaded microfibrils in tissues. Based on the microfibril morphologies observed after these experiments, on the crude collagenase cleavage sites identified in fibrillin-1, and on known antibody binding sites in fibrillin-1, a model is proposed in which fibrillin-1 molecules are staggered in microfibrils. This model further suggests that the N-terminal half of fibrillin-1 is asymmetrically exposed in the outer filaments, whereas the C-terminal half of fibrillin-1 is present in the interior of the microfibril.

UR - http://www.scopus.com/inward/record.url?scp=33947526687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33947526687&partnerID=8YFLogxK

U2 - 10.1074/jbc.M606370200

DO - 10.1074/jbc.M606370200

M3 - Article

C2 - 17158461

AN - SCOPUS:33947526687

VL - 282

SP - 4007

EP - 4020

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 6

ER -