Proteolytic Activity of IGFBP-3 in Various Clinical Conditions During Childhood Studied by Means of Western Immunoblotting

Yukihiro Hasegawa, Tomonobu Hasegawa, Hideko Koni, Taiji Aso, Noriko Tanaka, Shinobu Kotoh, Makoto Yamada, Yutaka Tsuchiya

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16 Citations (Scopus)


Insulin-like growth factors (IGFs) have 6 types of binding proteins (IGFBPs), and IGFBP-3 is the major IGFBP in human sera. A proteolytic enzyme for IGFBP-3 has recently been reported to be present in human and animal pregnant sera. Although the physiological significance of a pregnancy-associated IGFBP-3 protease remains to be established, the proteolysis could result in lowering the affinity for IGFs, thereby enhancing their delivery to target tissues by increasing free IGFs in the circulation. The methods for detection of IGFBP-3 protease which have been widely used so far are a method reported by Lamson et al. which used affinity crosslinking or western ligand blotting. These methods need radioactive materials (iodinated IGFs and IGFBP-3) and it takes at least a few days to get the results. We have now developed a simple assay for the proteolysis of IGFBP-3. The method is western immunoblotting without radioactive materials. The results can be obtained in a day. With this method, we proved the absence of significant proteolytic activity in sera from rapidly growing children (early stage of puberty or precocious puberty), and sera from a severe type of growth hormone deficiency. Significant proteolytic activity, as in pregnant women, was detected in 6 out of 11 patients with acute disorders such as measles, Kawasaki disease, bacterial meningitis and mycoplasma pneumonia, some of whom were probably in a catabolic condition. These data suggests that the proteolysis of IGFBP-3 might also be important in modulating IGF action in some acute diseases during childhood. The increased bioavailability of IGFs by IGFBP-3 proteolysis may play a role in overcoming catabolic conditions.

Original languageEnglish
Pages (from-to)569-576
Number of pages8
JournalEndocrine journal
Issue number4
Publication statusPublished - 1995
Externally publishedYes


  • IGFBP-3
  • IGFBP-3 Protease
  • Western immunoblotting

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology


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