Hydrophobic chromatography of fibronectin

Morgenthaler, J.‐J.

doi:10.1016/0014-5793(82)81308-2

Cited by 22 publications

(13 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the experiments described above we used fibronectin purified from plasma [17]. Although plasma and cellular fibronectins are very similar in molecular properties (they have nearly identical amino-acid compositions and secondary and tertiary structures), cellular fibronectin, in contrast to plasma fibronectin, is present not only in dimers but also in multimers [8].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Fibronectin-Cleaving Activity in Bronchial Secretions of Patients with Cystic Fibrosis

Suter

Schaad²,

Morgenthaler³

et al. 1988

Journal of Infectious Diseases

View full text Add to dashboard Cite

In cystic fibrosis, colonization of the airways with Pseudomonas aeruginosa follows colonization with Staphylococcus aureus and is related to accelerated deterioration of pulmonary function. Because P. aeruginosa adheres better to cell surfaces devoid of fibronectin, we searched for fibronectin-cleaving activity in bronchial secretions and saliva from 24 patients with cystic fibrosis who were followed up for 4.5 y and from two control groups. Proteolytic activity against 12sI-labeled fibronectin was continuously present in cysticfibrosis bronchial secretions; significantly higher fibronectin-cleaving activity was found in older vs. younger patients, in patients with advanced disease stages determined by a five-stage scoring system, and in those colonized with P. aeruginosa. The fibronectin-cleaving activity was due to neutrophil elastase and cathepsin G. Cystic fibrosis bronchial secretions had proteolytic activity against surface fibronectin of airway mucosal cells. Thus fibronectin-cleaving activity of bronchial secretions rather than of saliva may favor P. aeruginosa colonization of the upper respiratory tract in individuals with cystic fibrosis.Most individuals with cystic fibrosis who survive beyond the neonatal period die from respiratory failure, which is the consequence of chronic, progressively destructive bronchitis [1][2][3]. Clinical and pathological observations show extensive tissue destruction in the airways, which first causes obstruction and obliteration of small airways and thereafter destruction of the walls of large airways [4,5]. Early in the disease course the bacterial pathogens frequently found in the bronchial secretions are Staphylococcus aureus and Haemophilus influenzae. Later the airways of these subjects are invariably colonized with Pseudomonas aeruginosa, an event that initiates an acclerated deterioration of lung functions [3] as wellas an excessive systemic immune response [6]. These pathogens cannot be eliminated from the airways by antimicrobial therapy, and because the deleterious effect of P. aeruginosa infection in individuals with cystic fibrosis is well established, the identification of factors favoring colonization of the airways with P. aeruginosa is important. Coloniza-

show abstract

Section: Discussionmentioning

confidence: 99%

“…Fibronectin was purified from human plasma [17] and radiolabeled with 125 1 [18] (Amersham Corp., Amersham, England). About 250000 cpm of 125 1was incorporated into 1 ug of purified fibronectin.…”

Section: Specimens From All These Patients Bronchialmentioning

confidence: 99%

Fibronectin-Cleaving Activity in Bronchial Secretions of Patients with Cystic Fibrosis

Suter

Schaad²,

Morgenthaler³

et al. 1988

Journal of Infectious Diseases

View full text Add to dashboard Cite

show abstract

“…These sticky residues, at least some of which are also present in the synthetic III 1 peptide, could interact with other proteins or fragments in a nonspecific manner, in the same way for example that a number of Fn fragments have been shown to interact with alkyl-and aryl-Sepharose columns (17)(18)(19). In the present study, we used analytical affinity chromatography to test a complete set of fragments representing almost the entire light chain of Fn for their ability to bind native or denatured III 1 .…”

mentioning

confidence: 99%

Cryptic Self-association Sites in Type III Modules of Fibronectin

Ingham

Brew

Huff

et al. 1997

Journal of Biological Chemistry

109

View full text Add to dashboard Cite

In an effort to detect interactions between other Fn domains, all fragments were coupled to Sepharose, and each fragment was tested on each affinity matrix before and after denaturation. The only interaction detected was that of fluid phase III 1 with immobilized denatured 110-kDa CBF and 40-kDa Hep-2, both of which contain type III domains. Analysis of subfragments revealed this activity to be dominated by domains III 7 and III 15 . Fn itself did not bind to the denatured fragments. Thus, domain III 1 contains two cryptic "self-association sites," one that is buried in the core of the fold but recognizes many Fn fragments when presented as a peptide and another that is concealed in Fn but exposed in the native isolated domain and recognizes cryptic sites in two other type III domains. These interactions between type III domains could play an important role in assembly of Fn multimers in the extracellular matrix. Fibronectin (Fn)1 circulates in plasma as a 550-kDa 2-chain monomer that can be transformed by cultured fibroblasts into an insoluble fibrillar structure during the cell-driven process of matrix assembly (1). Several regions of Fn have been implicated in this process. N-terminal fragments containing the first five type I modules bind to cell layers and inhibit matrix assembly but are not themselves incorporated into the insoluble matrix unless bivalent, and then most efficiently in the presence of intact Fn (2-8). For a long time, it was thought that N-terminal fragments were interacting with a cell-surface "matrix assembly receptor" that was distinct from integrins that interact with Fn primarily via the Arg-Gly-Asp in the 10th type III domain. At least two unidentified molecules have been proposed as candidates for the matrix assembly receptor by virtue of their interaction with N-terminal fragments of Fn (9, 10). However, none has been further characterized or shown to play a role in matrix assembly. More recently, it is beginning to appear that the matrix assembly receptor, i.e. the molecule responsible for binding N-terminal fragments, might be Fn itself, perhaps conformationally altered by incorporation into the matrix (5, 11, 12).The fact that Fn matrix assembly could be inhibited with a monoclonal antibody directed to module III 1 (13) prompted efforts to examine the interaction of this module with Fn. Morla and Ruoslahti (14) showed that a synthetic 31-mer peptide with N-terminal sequence NAPQ . . . , derived from the middle of III 1 , was able to bind Fn, but the site of its interaction was not determined. The same group later reported that a longer recombinant peptide with the same N terminus, when incubated with whole Fn, was able to induce the formation of polymers that were stable in SDS in the absence of reducing agents and exhibited superior adhesive properties toward fibroblasts (15). At the same time, it was shown by Hocking et al.(11) that module III 1 , when adsorbed to plastic above its denaturation temperature, was able to bind Fn and its N-terminal 70-kDa fragment; other fragments...

show abstract

“…Fibronectin was quantitated by a laser nephelometric assay [14], Electrophoresis was car ried out in a Laemmli system [15] with a 7.5% poly acrylamide separating gel in the presence of sodium lauryl sulfate and 2-mercaptoethanol. The stained gels were scanned at 560 nm with a Cary 219 spectropho tometer equipped with a scanning attachment.…”

Section: Resultsmentioning

confidence: 99%