Baculovirus-dependent expression of membrane-associated and secreted, heavily modified proteins in insect cells often results in low yields. To optimize expression of T1, a heavily glycosylated receptor related to Interleukin-1 receptor type I, deletion mutants comprising either the heavily glycosylated extracellular domain or the isolated transmembrane and cytoplasmic portions of the T1 receptor were expressed from recombinant baculoviruses. As shown here, the use of a Baculovirus-derived leader sequence (GP67) in combination with an insect cell line with increased secretory capacity resulted in yields of several mg per liter (10(9) cells) of culture of both purified secretory T1 glycoprotein and membrane-associated T1 receptor.

Amino Acid Sequence↗

PubMed1995

Strain splitting of nitrogen acceptor levels in ZnSe.

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Journal Article↗

PubMed1995

Secreted and membrane-bound isoforms of T1, an orphan receptor related to IL-1-binding proteins, are differently expressed in vivo.

The murine T1 gene encodes a membrane-bound glycoprotein (T1-M), highly similar to interleukin-1 (IL-1) receptor type I, and a soluble variant (T1-S) representing its isolated extracellular domain. In vivo, the expression pattern of both T1 isoforms differs drastically. The T1-M receptor is abundantly expressed in single cells of the major hemopoietic organs (embryonic liver, spleen, bone marrow). It is restricted to few hemopoietic cell types throughout ontogenesis. By contrast, the soluble T1-S protein is predominantly expressed in selected nonhemopoietic embryonic tissues (developing skin, bone, and retina) and deposited in extracellular matrix. Despite the similarity of the T1 ligand-binding domain to all IL-1-binding proteins, it does not exhibit affinity to either IL-1 alpha or -beta. Thus, T1-M likely represents a novel orphan receptor of selected hemopoietic cells. The matrix-associated T1-S variant might act to create a reservoir of the putative T1 ligand in some differentiating tissues.

3T3 Cells↗

PubMed1995

Bone matrix deposition of T1, a homologue of interleukin 1 receptors.

The mouse T1 glycoprotein is a secreted molecule of the immunoglobulin superfamily with significant homology to interleukin 1 receptors. It is expressed during bone development, and the extracellular diffusible gene product is found associated with newly formed bone but not cartilage matrix. During osteogenic differentiation of mandibular condyles of newborn mice in vitro, T1 gene expression is induced shortly after cultivation and is observed throughout the differentiation process. The temporal expression pattern of the gene is an mandibular condyles indicates that T1 expression is an early marker of osteogenic differentiation. This view is substantiated by the analysis of T1 gene regulation in continuous osteogenic cell lines. Both in differentiating osteoblast-like KM-1K cells derived from mandibular condyles and in MC3T3 cells, T1 gene activity is preferentially associated with early differentiation stages. In mandibular condyles, the secreted extracellular T1 protein is deposited into newly formed osteoid but not into cartilage matrix. This novel bone matrix protein may locally modulate the availability of its ligand.

Animals↗

PubMed1994