The predominant form of secreted colony stimulating factor-1 is a proteoglycan

Laura K H Price, Haing U. Choi, Lawrence Rosenberg, E. Richard Stanley

Research output: Contribution to journalArticle

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Abstract

Colony stimulating factor-1 (CSF-1) is a homodimeric glycoprotein that humorally regulates the proliferation and differentiation of mononuclear phagocytic cells and locally regulates cells of the female reproductive tract. Alternative splicing of the human CSF-1 mRNA leads to alternative expression of the CSF-1 homodimer as a secreted glycoprotein or as a membrane-spanning molecule with cell surface biological activity. In the present study, analysis of immunoaffinity-purified CSF-1 from mouse L929 cell medium by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that CSF-1 is predominantly secreted as highly sulfated species of 375- and 250-kDa with a smaller amount of a 100-kDa species. Analysis by gel filtration in 4 M guanidine HC1 buffer, indicated that, in contrast to the 100-kDa species, the highly sulfated species exhibit anomalously high molecular weights and self-association on SDS-PAGE similar to the dermatan sulfate proteoglycan, biglycan. The three predominant CSF-1 species were shown to be an 80-kDa homodimer, an 80-kDa/50-kDa heterodimer, and a 50-kDa homodimer. The 80-kDa subunit contained a single 18-kDa chondroitin sulfate chain that was absent from the 50-kDa subunit. Furthermore, treatment of the 80- and 50-kDa subunits, synthesized in the presence of tunicamycin, with chondroitinase ABC, neuraminidase, and endo-́-N-acetyl galactosaminidase reduced their apparent molecular masses to 60 and 25 kDa, respectively. These results are consistent with intracellular proteolytic cleavage of the 80-kDa chondroitin sulfate containing subunits from the membrane spanning CSF-1 precursor at a point carboxyl-terminal to the single consensus sequence for glycosaminoglycan addition and cleavage of the 50-kDa glycoprotein subunit at a position aminoterminal to this site. The predominance of the proteoglycan form of secreted CSF-1, which represents only 3-4% of the total trichloroacetic acid-precipitable counts released from 35SO4 2-labeled L cells, has important implications for regulation by this growth factor.

Original languageEnglish (US)
Pages (from-to)2190-2199
Number of pages10
JournalJournal of Biological Chemistry
Volume267
Issue number4
StatePublished - Feb 5 1992

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Macrophage Colony-Stimulating Factor
Proteoglycans
Glycoproteins
Chondroitin Sulfates
Electrophoresis
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Biglycan
Chondroitin ABC Lyase
Membranes
Hexosaminidases
Tunicamycin
Trichloroacetic Acid
Guanidine
Consensus Sequence
Alternative Splicing
Neuraminidase
Molecular mass
Phagocytes
Bioactivity

ASJC Scopus subject areas

  • Biochemistry

Cite this

Price, L. K. H., Choi, H. U., Rosenberg, L., & Stanley, E. R. (1992). The predominant form of secreted colony stimulating factor-1 is a proteoglycan. Journal of Biological Chemistry, 267(4), 2190-2199.

The predominant form of secreted colony stimulating factor-1 is a proteoglycan. / Price, Laura K H; Choi, Haing U.; Rosenberg, Lawrence; Stanley, E. Richard.

In: Journal of Biological Chemistry, Vol. 267, No. 4, 05.02.1992, p. 2190-2199.

Research output: Contribution to journalArticle

Price, LKH, Choi, HU, Rosenberg, L & Stanley, ER 1992, 'The predominant form of secreted colony stimulating factor-1 is a proteoglycan', Journal of Biological Chemistry, vol. 267, no. 4, pp. 2190-2199.
Price LKH, Choi HU, Rosenberg L, Stanley ER. The predominant form of secreted colony stimulating factor-1 is a proteoglycan. Journal of Biological Chemistry. 1992 Feb 5;267(4):2190-2199.
Price, Laura K H ; Choi, Haing U. ; Rosenberg, Lawrence ; Stanley, E. Richard. / The predominant form of secreted colony stimulating factor-1 is a proteoglycan. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 4. pp. 2190-2199.
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