Characterization and partial purification of a keratinocyte-derived growth factor with wound-healing properties

✍ Scribed by Gunnar Kratz; Bror Jonzon; Anna Hultgård-Nilsson; Anders Hægerstrand

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 285 KB
• Volume: 15
• Category: Article
• ISSN: 0263-6484
• DOI: 10.1002/(sici)1099-0844(199709)15:3<153::aid-cbf734>3.0.co;2-c

✦ Synopsis

We have previously described the mitogenic and wound-healing properties of keratinocyte-conditioned medium (KCM). In this study we investigated the eect of KCM on the activation of second messenger systems and the expression of proto-oncogene in cultured human skin ®broblasts. We also present a partial puri®cation of the factor responsible for the mitogenic and wound-healing eects of KCM. KCM was shown to increase the expression of the proto-oncogenes c-fos, c-myc and c-jun. The eect of KCM on three second messenger systems was investigated. The extracellular release of choline metabolites was increased by 40 per cent when cells were stimulated with KCM whereas the formation of cAMP and hydrolysis of phosphatidyl inositol (PI) was unaected. KCM was puri®ed by ion exchange chromatography and ®ltration. The biologically active fraction was eluted from an SP column and retained its activity after ®ltration through a 3-kDa ®lter. The fraction was inactivated by heat and acid, indicative of a peptide origin. Furthermore, the active fraction was shown to increase the extracellular release of choline metabolites and to stimulate re-epithelialization in wounds in human skin in vitro comparable to KCM. The study indicates that human keratinocytes produce a 53 kDa peptide which may be partly responsible for the growth stimulatory and wound-healing properties of KCM.