Purification and structural analysis of a latent form of transforming growth factor-beta from rat platelets.
A latent form of transforming growth factor type-beta (TGF-beta) with a high molecular weight was purified to homogeneity from rat platelets by a six-step procedure. The yield of the purified latent TGF-beta from platelets of 2,500 rats was 1.4 mg. The purified latent TGF-beta was activated by treatment with urea at concentrations of over 4M or acidic solutions of below pH 4. SDS-PAGE and gel filtration chromatography showed that the latent TGF-beta consisted of active TGF-beta and glycoproteins of about 200 kDa as masking components, and that under physiological conditions, these components formed a high molecular weight complex of about 400 kDa linked by non-covalent bonds. Here, we found that the masking protein was composed of one large subunit of about 110 kDa and two small subunits of 39 kDa linked by disulfide bridges. The N-terminal amino acid sequence of the small subunit was identical to the N-terminal region of the TGF-beta precursor lacking a signal peptide. From these findings, we proposed a structural model for the latent TGF-beta from rat platelets.
Okada
F
F
Department of Biology, Faculty of Science, Kyushu University, Fukuoka.
Yamaguchi
K
K
Ichihara
A
A
Nakamura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001792:Blood Platelets / Q000032:analysis*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D015291:Transforming Growth Factors / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D000511:alpha-Macroglobulins / Q000378:metabolism
D007457:Iodine Radioisotopes; D000511:alpha-Macroglobulins; D015291:Transforming Growth Factors