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Purification and NH2-terminal sequence of a plasmacytoma growth factor derived from the murine macrophage cell line
Abstract:
Plasmacytoma growth factor (PCT-GF), a putative macrophage-derived lymphokine essential for the in vitro viability and proliferation of early generation plasmacytomas, was purified from conditioned medium of the murine macrophage cell line P388D1. The purification of PCT-GF was accomplished by a batch concentration on trimethylsilyl-controlled pore glass beads, followed by: gel filtration chromatography; hydrophobic interaction HPLC; and reverse-phase HPLC. SDS-PAGE analysis of the purified PCT-GF revealed a single band of Mr 23,000. The amino terminal sequence of PCT-GF was established as NH2-Pro-Thr-Ser-Gln-Val-Arg-Arg-Gly-Asp-Phe-Thr-Glu-Asp-Thr-Thr-Pro-Asn- Arg-Pro-Val-Tyr-Thr. No significant homology was found between this sequence and proteins in the National Biomedical Research Foundation database, suggesting that PCT-GF is a new lymphokine unrelated to previously described growth and differentiation factors.
Insights
Researchers purified plasmacytoma growth factor (PCT-GF), a macrophage-derived lymphokine crucial for plasmacytoma cell growth. This novel factor shows no homology to known growth factors, indicating a new class of lymphokine.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- Plasmacytoma growth factor (PCT-GF) is a macrophage-derived lymphokine.
- It is essential for the in vitro viability and proliferation of early plasmacytomas.
Purpose of the Study:
- To purify PCT-GF from conditioned medium of the murine macrophage cell line P388D1.
- To characterize the purified PCT-GF, including its molecular weight and N-terminal amino acid sequence.
Main Methods:
- Batch concentration using trimethylsilyl-controlled pore glass beads.
- Sequential chromatography: gel filtration, hydrophobic interaction HPLC, and reverse-phase HPLC.
- SDS-PAGE and amino acid sequencing.
Main Results:
- PCT-GF was purified to homogeneity.
- SDS-PAGE revealed a single band with a molecular weight of 23,000 (Mr 23,000).
- The N-terminal amino acid sequence was determined, showing no homology to known proteins in the NBRF database.
Conclusions:
- PCT-GF is a novel lymphokine.
- It is unrelated to previously described growth and differentiation factors.
- This finding suggests a new class of lymphokines involved in plasmacytoma biology.