Isolation of a heparan sulfate-containing proteoglycan from basement membrane.

Hassell, John R.; Robey, Pamela Gehron; Barrach, H. J.; Wilczek, Joseph; Rennard, Stephen I.; Martin, George R.

doi:10.1073/pnas.77.8.4494

Cited by 579 publications

(318 citation statements)

References 34 publications

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“…Similar molecular weights were observed for forms A and C on a molecular sieve column equilibrated in a low-ionic-strength buffer ( [6]. Perhaps remarkable is the moderately high cystine content (45 residues/1000) and low content of lysine (25 residues/lOOO).…”

Section: Size and Compositionsupporting

confidence: 54%

“…Our data characterize nidogen as a novel protein which differs from other basement membrane components such as laminin [4], collagen type IV [3,7], heparansulfate proteoglycan [6] and entactin [15,16] in its size, composition and immunochemical properties. Nidogen consists of a single polypeptide chain with M, = 80000.…”

Section: Discussionmentioning

confidence: 99%

“…The EHS sarcoma was used for purification of acidextracted collagen type IV [20], laminin [4], laminin fragments [22] and proteoglycan BM-1 [6]. The latter component was extracted with neutral salt (see above) and purified by chromatography on DEAE-cellulose at pH 4.0 and on Sephacryl S-400 followed by CsCl gradient centrifugation.…”

Section: Preparation Of Other Basement Membrane Proteinsmentioning

confidence: 99%

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Nidogen: a new, self‐aggregating basement membrane protein

Timpl

Dziadek

Fujiwara

et al. 1983

European Journal of Biochemistry

253

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Nidogen was purified from a mouse tumor basement membrane where it accounted for 2 -3 of the total proteins. It was isolated as two forms (A and B) of a monomer (Mr = 80000) each consisting of a single polypeptide chain folded into a globular head connected to a small tail. The B form of the monomer was shown to be capable of aggregating into a nest-like structure (Mr > 250000). A smaller form (M, = 45000) was observed in some of the extracts. The amino acid composition of nidogen was different to that of other basement membrane proteins. It contained about 10 % carbohydrate, with N-linked and 0-linked oligosaccharide chains in similar proportions.Isoelectrofocussing demonstrated a limited heterogeneity of nidogen with PI in the range 6.5 -7. Monomeric nidogen failed to interact with other basement membrane components and heparin. Aggregation could be induced by limited proteolysis and was reversed by detergents or high salt concentrations. Together with the observation that most of the nidogen could be solubilized only after destroying the collagenous matrix, the data indicate that aggregation of nidogen reflects an activity involved in matrix assembly. Specific antibodies raised against nidogen did not distinguish between the monomeric and aggregated form of the protein but showed that the fragment was antigenically deficient. These antibodies did not cross-react with collagen type IV, laminin, entactin and heparansulfate proteoglycan. Immunofluorescence staining and absorption studies demonstrated that nidogen is a common component of authentic basement membranes. Larger forms of nidogen (Mr about 100000 and 150000) were found in organ cultures of Reichert's membrane suggesting that it is synthesized in precursor forms.Basement membranes are extracellular sheets of proteins in close vicinity to epithelial, endothelial, muscle and other cells of the body. They separate these cells from underlying connective tissue and serve as major filtration barriers and for maintaining a proper tissue organization [1,2]. Major components of basement membranes are collagen type IV [3], laminin [4] and heparansulfate proteoglycan [5, 61. The collagenous proteins are responsible for the mechanical stability of these matrices and are integrated in a kind of a network using their terminal domains for self-assembly [7]. Laminin and proteoglycan have the ability to interact with each other and presumably mediate binding of cells to the network [2,8,9]. The precise architecture of these extracellular structures has not yet been elucidated. The difficulties of structural studies are due in part to the complexity and extraordinary size (length 100 -400 nm) of the known components, but also to the incomplete knowledge of additional constituents which, although minor by weight, may nevertheless be important for the structural integrity and function of basement membranes.A considerable diversity of basement membrane components was indicated in previous immunological studies [3,10] and in electrophoretic analyses of glomerular and other...

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Section: Size and Compositionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Other Basement Membrane Proteinsmentioning

confidence: 99%

See 1 more Smart Citation

Nidogen: a new, self‐aggregating basement membrane protein

Timpl

Dziadek

Fujiwara

et al. 1983

European Journal of Biochemistry

253

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“…Five monoclonal antibodies that recognize determinants on HeSPG and precipitate the neuritepromoting activity in CM, bind to the surface of these cells in culture (33). These DRG-derived cells were incubated with 35S-labeled sulfate, a procedure that primarily labels proteoglycans (22,35). The INO antigen was coeluted from DEAE cellulose between 0.3 and 0.5 M NaCI and chromatographed on Sepharose 4B.…”

Section: Composition Of the Ino-binding Sitementioning

confidence: 99%

A monoclonal antibody that blocks the activity of a neurite regeneration-promoting factor: studies on the binding site and its localization in vivo.

Chiu¹,

Matthew²,

Patterson³

1986

The Journal of Cell Biology

111

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Abstract. Work from several laboratories has identified a proteoglyean complex secreted by a variety of non-neuronal cells that can promote neurite regeneration when applied to the surface of culture dishes. Using a novel immunization protocol, a monoclonal antibody (INO) was produced that blocks the activity of this outgrowth-promoting factor (Matthew, W. D., and P. H. Patterson, 1983, Cold Spring Harbor Symp. Quant. Biol. 48:625-631). We have used the antibody to analyze the components of the active site and to localize the complex in vivo. INO binding is lost when the complex is dissociated; if its components are selectively reassociated, INO binds only to a complex containing two different molecular weight species. These are likely to be laminin and heparan sulfate proteoglycan, respectively. On frozen sections of adult rat tissues, INO binding is present on the surfaces of glial cells of the peripheral, but not the central, nervous system. INO also binds to the basement membrane surrounding cardiac and skeletal muscle cells, and binding to the latter greatly increases after denervation. In the adrenal gland and kidney, INO selectively reacts with areas rich in basement membranes, staining a subset of structures that are immunoreactive for both laminin and heparan sulfate proteoglycan. In general, the outgrowth-blocking antibody binds to areas known to promote axonal regeneration and is absent from areas known to lack this ability. This suggests that this complex, which is active in culture, may be the physiological substrate supporting nerve regeneration in vivo.oNs damaged in the peripheral nervous system (PNS) 1 of adult mammals have the capacity to regenerate over long distances. The prerequisites for successful neurite outgrowth include the presence of trophic factors, such as nerve growth factor (3), and/or an appropriate surface on which to grow (30). The molecular nature of the surfaces that allow or stimulate regeneration in vivo is not known. Experiments in culture, however, have delineated a number of candidate molecules that can promote neurite outgrowth under specific conditions. For instance, surfaces coated with laminin or fibronectin stimulate a much more rapid and extensive regeneration of neuronal processes than can serum-or collagen-coated plastic (2,4,9,24,29,31,39,44). The finding that laminin, in particular, provides an especially good substrate for outgrowth is of interest because a variety of non-neuronal cells secrete neurite regeneration-promoting factors (1,12,14,26), some of which are extracellular matrix complexes containing primarily laminin and heparan sulfate proteoglycan (HeSPG) (8,16,28).Since laminin is a component of a naturally occurring,1. Abbreviations used in this paper: CM, conditioned medium; CNS, central nervous system; DRG, dorsal root ganglion; HeSPG, heparan sulfate proteoglycan; PNS, peripheral nervous system. neurite-promoting complex, and purified laminin can stimulate outgrowth in culture, the question arises as to whether there is a role for the HeSPG compo...

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“…Several protein components of basement membranes have been identified (4). Type IV collagen appears to be restricted to the lamina densa (5-7), whereas laminin, fibronectin, and heparan sulfate proteoglycan are present in either the lamina lucida or lamina densa (5,(7)(8)(9)(10)(11)(12). We have shown that antibodies to type IV collagen, laminin, and the thymocyte differentiation antigen Thy-1 stain the basement membrane in the mammary glands of both virgin and lactating rats at the light microscope level, whereas antibodies to fibronectin only stain the basement membrane in the mammary gland of lactating rats (1,13,14).…”

mentioning

confidence: 99%

Topographical arrangement of basement membrane proteins in lactating rat mammary gland: comparison of the distribution of type IV collagen, laminin, fibronectin, and Thy-1 at the ultrastructural level.

Monaghan¹,

Warburton²,

Perusinghe³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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The topographical distribution of type IV collagen, laminin, fibronectin, and the thymocyte differentiation antigen Thy-i in the basement membrane of the lactating rat mammary gland was investigated. Small cubes of tissue, which had not been subjected to prior fixation or freezing, were incubated with monospecific or monoclonal antibodies to these proteins, and the antibodies were located by an indirect immunoperoxidase staining technique and observed in the electron microscope. The lamina densa stained uniformly with antibodies to type IV collagen and laminin. In addition, both laminin and type IV collagen were present in semiperiodic clusters that traversed the lamina lucida from the cell surface to the lamina densa. Fibronectin was present only in the semiperiodic clusters and not elsewhere in the basement membrane. These clusters were irregularly spaced along the cell surface and heterogeneous in size. It remains to be determined if these three proteins are present in the same clusters. Thy-I was largely present on the lamina densa and not on the lamina lucida.The Thy-i staining of the lamina densa occurred in discrete maxima and minima. These maxima occurred in regions adjacent to Thy-l-bearing stromal cells. Thus, the topographical distribution of proteins within a basement membrane varies in a nonrandom manner, and local factors can modify this distribution. We suggest that this topographical variability may play a role in cell recognition and signalling processes that occur across the basement membrane.In the rodent and human mammary glands, the parenchyma are separated from the surrounding connective tissue by a basement membrane (1). Ultrastructurally, this basement membrane consists of an electron-lucent region, the lamina lucida, directly adject to the epithelial and myoepithelial cells, and an electron dense region, the lamina densa, located between the lamina lucida and the connective tissue (2). In some tissues anchoring fibrils that extend from the lamina densa into the surrounding connective tissue have been described (3). Several protein components of basement membranes have been identified (4). Type IV collagen appears to be restricted to the lamina densa (5-7), whereas laminin, fibronectin, and heparan sulfate proteoglycan are present in either the lamina lucida or lamina densa (5, 7-12). We have shown that antibodies to type IV collagen, laminin, and the thymocyte differentiation antigen Thy-1 stain the basement membrane in the mammary glands of both virgin and lactating rats at the light microscope level, whereas antibodies to fibronectin only stain the basement membrane in the mammary gland of lactating rats (1,13,14). We now report on the immunocytochemical distribution at the ultrastructural level of type IV collagen, laminin, fibronectin, and Thy-i in the basement membrane of the lactating rat mammary gland. METHODSAntibodies. Monospecific antibodies to type IV collagen and laminin, both purified from the mouse Engelbreth-Holm-Swarm (EHS) sarcoma, and rat plasma fibronectin hav...

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Isolation of a heparan sulfate-containing proteoglycan from basement membrane.

Cited by 579 publications

References 34 publications

Nidogen: a new, self‐aggregating basement membrane protein

Nidogen: a new, self‐aggregating basement membrane protein

A monoclonal antibody that blocks the activity of a neurite regeneration-promoting factor: studies on the binding site and its localization in vivo.

Topographical arrangement of basement membrane proteins in lactating rat mammary gland: comparison of the distribution of type IV collagen, laminin, fibronectin, and Thy-1 at the ultrastructural level.

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