Alveolar Basement Membrane: Molecular Properties of the Noncollagenous Domain (Hexamer) of Collagen IV and its Reactivity with Goodpasture Autoantibodies

Gunwar, Sripad; Bejarano, P.A.; Kalluri, Raghu; Langeveld, J.P.M.; Wisdom, Billie J.; Noelken, Milton E.; Hudson, Billy G.

doi:10.1165/ajrcmb/5.2.107

Cited by 47 publications

(44 citation statements)

References 14 publications

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“…Truncated type IV collagen protomers were produced by digestion of BMs with pseudolysin (Pseudomonas aeruginosa elastase; EC 3.4.24.26; purchased from Nagase Biomedical, Fukushiyama, Japan) (14). Aorta BM (80 g of wet ground tissue) was digested with 0.5% pseudolysin for 24 h at 4°C, then the insoluble fraction was further digested at 25°C for 24 h as described previously (15). The reaction was arrested by the addition of 20 mM EDTA, and then the truncated protomers solubilized by pseudolysin digestion were twice precipitated with 2 M NaCl and resolubilized in Tris-buffered saline (0.15 M NaCl and 50 mM Tris-HCl, pH 7.5) for electron microscopy.…”

Section: Methodsmentioning

confidence: 99%

The NC1 Domain of Collagen IV Encodes a Novel Network Composed of the α1, α2, α5, and α6 Chains in Smooth Muscle Basement Membranes

Borza¹,

Bondar²,

Ninomiya³

et al. 2001

Journal of Biological Chemistry

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133

119

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Type IV collagen, the major component of basement membranes (BMs), is a family of six homologous chains (␣1-␣6) that have a tissue-specific distribution. The chains assemble into supramolecular networks that differ in the chain composition. In this study, a novel network was identified and characterized in the smooth muscle BMs of aorta and bladder. The noncollagenous (NC1) hexamers solubilized by collagenase digestion were fractionated by affinity chromatography using monoclonal antibodies against the ␣5 and ␣6 NC1 domains and then characterized by two-dimensional gel electrophoresis and Western blotting. Both BMs were found to contain a novel ␣1⅐␣2⅐␣5⅐␣6 network besides the classical ␣1⅐␣2 network. The ␣1⅐␣2⅐␣5⅐␣6 network represents a new arrangement in which a protomer (triplehelical isoform) containing the ␣5 and ␣6 chains is linked through NC1-NC1 interactions to an adjoining protomer composed of the ␣1 and ␣2 chains. Re-association studies revealed that the NC1 domains contain recognition sequences sufficient to encode the assembly of both networks. These findings, together with previous ones, indicate that the six chains of type IV collagen are distributed in three major networks (␣1⅐␣2, ␣3⅐␣4⅐␣5, and ␣1⅐␣2⅐␣5⅐␣6) whose chain composition is encoded by the NC1 domains. The existence of the ␣1⅐␣2⅐␣5⅐␣6 network provides a molecular explanation for the concomitant loss of ␣5 and ␣6 chains from the BMs of patients with X-linked Alport's syndrome. The basement membrane (BM),1 a continuous sheet of extracellular matrix, separates epithelial cells from the underlying stroma and plays important roles in normal biological functions (such as cell adhesion, growth, and differentiation; tissue repair; and molecular ultrafiltration) as well as in pathological events (such as cancer cell invasion and metastasis). Moreover, degradation and de novo synthesis of vascular BMs are critical events in the angiogenesis processes. BMs function is impaired in hereditary and acquired diseases in which type IV collagen is affected, including Alport's syndrome, a hereditary form of progressive renal disease; diffuse leiomyomatosis, a benign proliferation of smooth muscle cells; and Goodpasture syndrome, an anti-type IV collagen autoimmune disease (1).Type IV collagen is the major structural component of the BM, and it consists of a family of six homologous ␣(IV) chains, designated ␣1-␣6 (1). Each chain is characterized by a long collagenous domain of ϳ1400 residues of Gly-X-Y repeats, interrupted by ϳ20 short noncollagenous sequences, and by a noncollagenous (NC1) domain of ϳ230 residues at the carboxyl terminus. Three ␣(IV) chains assemble into triple-helical molecules (protomers) that further associate to form supramolecular networks by dimerization at the carboxyl terminus through NC1 domains and by formation of tetramers at the amino terminus (2). The chain composition, and thus the properties of the type IV collagen networks are influenced by two factors. First, the chain composition of networks is limited by chain availability...

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Section: Methodsmentioning

confidence: 99%

The NC1 Domain of Collagen IV Encodes a Novel Network Composed of the α1, α2, α5, and α6 Chains in Smooth Muscle Basement Membranes

Borza¹,

Bondar²,

Ninomiya³

et al. 2001

Journal of Biological Chemistry

Self Cite

133

119

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“…Rabbit antibodies were prepared and characterized to all six ␣ -chains of type IV collagen as recombinant molecules (7,25,26). Alport alloantibodies and Goodpasture autoantibodies were also described previously (7,27).…”

Section: Methodsmentioning

confidence: 99%

“…The suspension was centrifuged, washed three times with ice cold distilled water, each time homogenizing with a polytron homogenizer (28). To solubilize the noncollagenous domain of collagen IV, renal basement membrane preparations were digested with bacterial collagenase at 37 Њ C for 24 h in the following digestion buffer: 0.05 M Hepes, pH 7.5, 0.01 M CaCl 2 , 4 mM N -ethylmaleimide, 5 mM benzamidine HCl, 1 mM phenylmethanesulfonyl fluoride, and 25 mM e-aminohexanoic acid (7,29).…”

Section: Methodsmentioning

confidence: 99%

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Isoform switching of type IV collagen is developmentally arrested in X-linked Alport syndrome leading to increased susceptibility of renal basement membranes to endoproteolysis.

Kalluri¹,

Shield²,

Todd³

et al. 1997

J. Clin. Invest.

283

230

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Normal glomerular capillaries filter plasma through a basement membrane (GBM) rich in ␣ 3(IV), ␣ 4(IV), and ␣ 5(IV) chains of type IV collagen. We now show that these latter isoforms are absent biochemically from the glomeruli in patients with X-linked Alport syndrome (XAS). Their GBM instead retain a fetal distribution of ␣ 1(IV) and ␣ 2(IV) isoforms because they fail to developmentally switch their ␣ -chain use. The anomalous persistence of these fetal isoforms of type IV collagen in the GBM in XAS also confers an unexpected increase in susceptibility to proteolytic attack by collagenases and cathepsins. The incorporation of cysteine-rich ␣ 3(IV), ␣ 4(IV), and ␣ 5(IV) chains into specialized basement membranes like the GBM may have normally evolved to protectively enhance their resistance to proteolytic degradation at the site of glomerular filtration. The relative absence of these potentially protective collagen IV isoforms in GBM from XAS may explain the progressive basement membrane splitting and increased damage as these kidneys deteriorate. ( J. Clin. Invest. 1997. 99:2470-2478.)

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“…The reactions were terminated by boiling with sample buffer and the proteins electrophoretically separated by SDS-PAGE. Direct ELISA As previously described (Kalluri et al, 1997), type IV collagen from Calbiochem (San Diego, CA) or type IV collagen protomer and individual domains (7 S, triple helical and NC1), isolated from bovine kidney (Gunwar et al, 1991;Kalluri et al, 1996) were used to coat ELISA plates at a concentration of 200 ng per well and blocked with 2% BSA in PBS. Usherin domains (Ts, FN, LN and LE fusion proteins) were incubated with each type IV collagen domain and usherin domain binding was probed using rabbit anti-GST antibodies and sheep anti-rabbit IgG secondary antibodies conjugated to alkaline phosphatase.…”

Section: Electrophoresis Of Protein Under Denaturing or Nondenaturingmentioning

confidence: 99%

A domain-specific usherin/collagen IV interaction may be required for stable integration into the basement membrane superstructure

Bhattacharya

Kalluri

Orten

et al. 2004

Journal of Cell Science

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Usherin is a basement membrane protein encoded by the gene associated with Usher syndrome type IIa, the most common deaf/blind disorder. This report demonstrates a specific interaction between type IV collagen and usherin in the basement membrane, with a 1:1 stoichiometry for binding. Genetic and biochemical approaches were used to explore the role of type IV collagen binding in usherin function. We demonstrate binding occurs between the LE domain of usherin and the 7S domain of type IV collagen. A purified fusion peptide comprising the first four LE modules was shown to compete with full-length recombinant usherin for type IV collagen binding. However, synonymous fusion peptides with single amino acid substitutions resulting from missense mutations that were known to cause Usher syndrome type IIa in humans, failed to compete. Only mutations in loop b of the LE domain abolished binding activity. Co-immunoprecipitation and western blot analysis of testicular basement membranes from the Alport mouse model show a 70% reduction in type IV collagen is associated with a similar reduction in usherin, suggesting the usherin/collagen (IV) interaction stabilizes usherin in the basement membrane. Thus, the domain-specific interaction between usherin and type IV collagen appears essential to usherin stability in vivo, and loss of this interaction may result in Usher pathology in humans.

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Alveolar Basement Membrane: Molecular Properties of the Noncollagenous Domain (Hexamer) of Collagen IV and its Reactivity with Goodpasture Autoantibodies

Cited by 47 publications

References 14 publications

The NC1 Domain of Collagen IV Encodes a Novel Network Composed of the α1, α2, α5, and α6 Chains in Smooth Muscle Basement Membranes

The NC1 Domain of Collagen IV Encodes a Novel Network Composed of the α1, α2, α5, and α6 Chains in Smooth Muscle Basement Membranes

Isoform switching of type IV collagen is developmentally arrested in X-linked Alport syndrome leading to increased susceptibility of renal basement membranes to endoproteolysis.

A domain-specific usherin/collagen IV interaction may be required for stable integration into the basement membrane superstructure

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