Formation of collagen fibrils in vitro by cleavage of procollagen with procollagen proteinases.

Miyahara, Michinori; Njieha, F K; Prockop, Darwin J.

doi:10.1016/s0021-9258(18)34351-5

Cited by 119 publications

(13 citation statements)

References 25 publications

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“…This threshold proved to be 2 g/ml for Vitrogen collagen and 5 g/ml for rat tail collagen. At higher concentrations, fibrils 6 to 12 m in length and similar to self-assembled fibrils previously described 7,8 settled onto the fibronectincoated coverslips (Figure 1a). At concentrations below these thresholds, fibrils were not detected on the substrate or suspended in the media (Figure 1b).…”

Section: Smc-associated Collagen Fibril Assembly Is Distinct From Cell-free Fibril Assemblysupporting

confidence: 78%

“…This action reduces the solubility of the protein and initiates the entropy-driven self-assembly process. 8,9 Although only the latter approach involves the physiologically relevant step of procollagen cleavage, both in vitro systems yield early collagen fibrils with characteristics similar to those found in developing tissues. 3,10 Moreover, both approaches have been valuable for elucidating conditions for collagen self-assembly and in defining controlling elements for this within the collagen molecule.…”

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confidence: 99%

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Vascular Smooth Muscle Cells Orchestrate the Assembly of Type I Collagen via α2β1 Integrin, RhoA, and Fibronectin Polymerization

Diepstraten

D’Souza

et al. 2003

The American Journal of Pathology

151

143

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Assembly of collagen into fibrils is widely studied as a spontaneous and entropy-driven process. To determine whether vascular smooth muscle cells (SMCs) impact the formation of collagen fibrils, we microscopically tracked the conversion of soluble to insoluble collagen in human SMC cultures, using fluorescent type I collagen at concentrations less than that which supported self-assembly. Collagen microaggregates were found to form on the cell surface, initially as punctate collections and then as an increasingly intricate network of fibrils. These fibrils displayed 67-nm periodicity and were found in membrane-delimited cellular invaginations. Fibril assembly was inhibited by an anti-alpha2beta1 integrin antibody and accelerated by an alpha2beta1 integrin antibody that stimulates a high-affinity binding state. Newly assembled collagen fibrils were also found to co-localize with newly assembled fibronectin fibrils. Moreover, inhibition of fibronectin assembly with an anti-alpha5beta1 integrin antibody completely inhibited collagen assembly. Collagen fibril formation was also linked to the cytoskeleton. Fibrils formed on the stretched tails of SMCs, ran parallel to actin microfilament bundles, and formed poorly on SMCs transduced with retrovirus containing cDNA for dominant-negative RhoA and robustly on SMCs expressing constitutively active RhoA. Lysophosphatidic acid, which activates RhoA and stimulates fibronectin assembly, stimulated collagen fibril formation, establishing for the first time that collagen polymerization can be regulated by soluble agonists of cell function. Thus, collagen fibril formation is under close cellular control and is dynamically integrated with fibronectin assembly, opening new possibilities for modifying collagen deposition.

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Section: Smc-associated Collagen Fibril Assembly Is Distinct From Cell-free Fibril Assemblysupporting

confidence: 78%

mentioning

confidence: 99%

Vascular Smooth Muscle Cells Orchestrate the Assembly of Type I Collagen via α2β1 Integrin, RhoA, and Fibronectin Polymerization

Diepstraten

D’Souza

et al. 2003

The American Journal of Pathology

151

143

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“…These results have led to the suggestion that the amino-terminal propeptide may play a role in longitudinal fibril assembly and provide a mechanism of controlling lateral fibril growth (13,14). Removal of the carboxyl propeptide has also been shown to be a prerequisite for fibril formation in vitro (32). The data presented here provide a direct measurement by pulse-chase analysis of processing kinetics versus collagen incorporation into an extracellular matrix and conclusively demonstrate the relationship between matrix maturation and the rate of procollagen processing.…”

Section: Discussionmentioning

confidence: 99%

Collagen expression, ultrastructural assembly, and mineralization in cultures of chicken embryo osteoblasts.

Gerstenfeld

Chipman

Kelly

et al. 1988

The Journal of Cell Biology

164

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Abstract. A newly defined chick calvariae osteoblast culture system that undergoes a temporal sequence of differentiation of the osteoblast phenotype with subsequent mineralization (Gerstenfeld, L. C., S. Chipman, J. Glowacki, and J. B. Lian. 1987. Dev. Biol. 122:49-60) has been examined for the regulation of collagen synthesis, ultrastructural organization of collagen fibrils, and extracellular matrix mineralization. Collagen gene expression, protein synthesis, processing, and accumulation were studied in this system over a 30-d period. Steady state mRNA levels forpro al(I) and pro ~t2 collagen and total collagen synthesis increased 1.2-and 1.8-fold, respectively, between days 3 and 12. Thereafter, total collagen synthesis decreased 10-fold while mRNA levels decreased 2.5-fold. In contrast to the decreasing protein synthesis after day 12, total accumulated collagen in the cell layers increased sixfold from day 12 to 30. Examination of the kinetics of procollagen processing demonstrated that there was a sixfold increase in the rate of procollagen conversion to ~t chains from days 3 to 30 and the newly synthesized collagen was more efficiently incorporated into the extracellular matrix at later culture times.The macrostructural assembly of collagen and its relationship to culture mineralization were also examined. High voltage electron microscopy demonstrated that culture cell layers were three to four cells thick. Each cell layer was associated with a layer of well developed collagen fibrils orthogonally arranged with respect to adjacent layers. Fibrils had distinct 64-70-nm periodicity typical of type I collagen. Electron opaque areas found principally associated with the deepest layers of the fibrils consisted of calcium and phosphorus determined by electron probe microanalysis and were identified by electron diffraction as a very poorly crystalline hydroxyapatite mineral phase.These data demonstrate for the first time that cultured osteoblasts are capable of assembling their collagen fibrils into a bone-specific macrostructure which mineralizes in a manner similar to that characterized in vivo. Further, this matrix maturation may influence the processing kinetics of the collagen molecule.T HE processes by which mineralization is initiated and regulated in bone and other vertebrate calcifying tissues are incompletely understood. It is known, however, that a prerequisite for mineralization is the synthesis and assembly of an extracellular matrix into which mineral may be deposited. Collagen type I has been shown to be the major extracellular matrix protein of bone. It comprises between 60-70% of its organic components and between 20-30% of its total dry mass (20). Physiologically, type I collagen provides the protein basis for the architecture of bone and the scaffold into which mineral is accumulated (5,20,41). The importance of collagen type I in maintaining structural integrity and proper mineralization of bone has been demonstrated for one form of inherited osteogenesis imperfecta. A specific frame shift m...

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“…Fibroblasts regulate the stoichiometry and sequence of mixing of the extracellular matrix components within the cell and influence matrix formation through the vectoral discharge of these packaged matrix components into the extracellular space (15,16). Postdepositional enzymatic modifications, such as procollagen processing and covalent cross-linking, are also important in the establishment of matrix order (17)(18)(19)(20) and must be regulated by the fibroblasts (21). In embryonic tendons it has been demonstrated that fibril formation occurs in intimate association with the fibroblast cell surface (22).…”

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confidence: 99%

Extracellular compartments in matrix morphogenesis: collagen fibril, bundle, and lamellar formation by corneal fibroblasts.

Birk¹,

Trelstad²

1984

The Journal of Cell Biology

266

172

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The regulation of collagen fibril, bundle, and lamella formation by the corneal fibroblasts, as well as the organization of these elements into an orthogonal stroma, was studied by transmission electron microscopy and high voltage electron microscopy. Transmission and high voltage electron microscopy of chick embryo corneas each demonstrated a series of unique extracellular compartments. Collagen fibrillogenesis occurred within small surface recesses. These small recesses usually contained between 5 and 12 collagen fibrils with typically mature diameters and constant intrafibrillar spacing. The lateral fusion of the recesses resulted in larger recesses and consequent formation of prominent cell surface foldings. Within these surface foldings, bundles that contained 50-100 collagen fibrils were formed. The surface foldings continued to fuse and the cell surface retracted, forming large surface-associated compartments in which bundles coalesced to form lamellae. High voltage electron microscopy of 0.5 p,m sections cut parallel to the corneal surface revealed that the corneal fibroblasts and their processes had two major axes at approximately right angles to one another. The surface compartments involved in the production of the corneal stroma were aligned along the fibroblast axes and the orthogonality of the cell was in register with that of the extracellular matrix. In this manner, corneal fibroblasts formed collagen fibrils, bundles, and lamellae within a controlled environment and thereby determined the architecture of the corneal stroma by the configuration of the cell and its associated compartments.The rigid control of collagen fibril structure and the arrangement of fibrils into a specific three-dimensional architecture is necessary for the development of a transparent corneal stroma. In this paper we demonstrate how the corneal fibroblast exerts control over collagen fibrillogenesis and the positioning of newly formed fibrils into a highly ordered lamellar matrix.The mature chick corneal stroma is composed of collagen fibrils arranged as lamellae parallel to the corneal surface. Collagen fibrils within a lamella have the same orientation; collagen fibrils in adjacent layers are oriented approximately at fight angles, forming an orthogonal grid. The orthogonal corneal lamellae are composed of small diameter (25 nm) collagen fibrils equidistantly spaced (1, 2). The orthogonal lamellae describe a gradual clockwise shift from epithelium to endothelium of approximately 220 °. This clockwise spiral pattern resembles a cholesteric liquid crystal and has the same handedness in both eyes (2, 3); in contrast, all other ocular features, such as the pattern of overlap of the scleral ossicles, demonstrate mirror symmetry (4).Morphogenesis of the chick corneal stroma occurs in a sequence of relatively well-described stages (for review, see reference 5). Initially, the corneal epithelium deposits the primary corneal stroma beneath its basal surface. The pattern of this epithelially derived acellular stroma is i...

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Formation of collagen fibrils in vitro by cleavage of procollagen with procollagen proteinases.

Cited by 119 publications

References 25 publications

Vascular Smooth Muscle Cells Orchestrate the Assembly of Type I Collagen via α2β1 Integrin, RhoA, and Fibronectin Polymerization

Vascular Smooth Muscle Cells Orchestrate the Assembly of Type I Collagen via α2β1 Integrin, RhoA, and Fibronectin Polymerization

Collagen expression, ultrastructural assembly, and mineralization in cultures of chicken embryo osteoblasts.

Extracellular compartments in matrix morphogenesis: collagen fibril, bundle, and lamellar formation by corneal fibroblasts.

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