TGF-beta binding in human Wharton's jelly.
Our previous study reported that TGF-beta may be isolated from human Wharton's jelly (WJ) in a form of soluble, high molecular complex(es). We decided to study the effect of extracellular matrix degradation and reduction of disulphide bridges reduction on the release of TGF-beta from WJ. The WJ prepared from the umbilical cords of newborns delivered at term by healthy mothers was homogenised and treated with hyaluronidase, collagenase, heparinase, chondroitinase and beta-mercaptoethanol, the resulting extracts were then submitted to TGF-beta immunoassay and SDS/PAGE followed by Western immunoblotting. The effect of metalloproteinase activation on TGF-beta was also studied. Pre-treatment of WJ homogenates with hyaluronidase or collagenase markedly increased the extractability of TGF-beta, but did not dissociate the complexes. In contrast, the action of beta-mercaptoethanol resulted in the release of free TGF-beta; but activation of metalloproteinases resulted in the disappearance of this factor. We conclude that TGF-beta1 is bound through disulphide bonds to an extracellular matrix component of WJ. The large amount of collagen fibrils and hyaluronate molecules which surround the cells scattered in WJ may prevent the access of extracting solution to TGF-beta causing a low extractability of this factor. Although hyaluronate and collagen do not bind TGF-beta directly, they may present a barrier that prevents the diffusion of TGF-beta in WJ and results in its concentration around the cells thereby facilitating its interaction with membrane receptors and subsequent stimulation of cell division and synthesis of extracellular matrix components.
Małkowski
Andrzej
A
Department of Medical Biochemistry, Medical Academy of Bialystok, ul. A. Mickiewicza 2C, 15-089 Bialystok, Poland. amalkow@amb.edu.pl
Sobolewski
Krzysztof
K
Jaworski
Stefan
S
Bańkowski
Edward
E
D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D020782:Matrix Metalloproteinases / Q000378:metabolism; D011247:Pregnancy; D011485:Protein Binding; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014470:Umbilical Cord / Q000033:anatomy & histology / Q000737:chemistry / Q000378:metabolism
D016212:Transforming Growth Factor beta; D020782:Matrix Metalloproteinases