Tetranectin, a plasminogen kringle 4-binding protein. Cloning and gene expression pattern in human colon cancer

Tetranectin is a recently discovered protein that binds to kringle 4 region of plasminogen (Clemmensen I, Petersen LC, Kluft C. Eur J Biochem 1986; 156:327.

The mRNA encoding human tetranectin was cloned by using degenerate primers in a reverse transcriptase reaction followed by polymerase chain reaction amplification. The resulting polymerase chain reaction product was examined by DNA sequencing and subsequently used as probe for screening a human placental cDNA library. A full length cDNA clone (TET-1) was isolated, characterized, and used for Northern blot and in situ hybridization.

DNA sequencing analysis revealed a 874-base pair cDNA containing an open reading frame of 606 base pairs encoding 202 amino acids. A classical signal peptide was present starting with the initiation methionine. The mature tetranectin chain consisted of 181 amino acids (M(r) = 20,169). The 3' noncoding region contained a single polyadenylation signal and a 26-residue poly A tail. The predicted amino acid sequence of the mature tetranectin chain showed, except for one amino acid, complete identity to that obtained by sequencing of the native protein (Fuhlendorff J, Clemmensen I, Magnusson S, Biochemistry 1987;26:6757). Northern blot of poly A+ revealed a single band of approximately 1 kb. Northern blot analysis of poly A+ isolated from a series of normal human tissues (lung, liver, spleen, kidney, and pancreas) revealed a distinct hybridization band that was especially prominent in the lungs and spleen. No hybridization signal was detected in three carcinoma cell lines examined in parallel. Northern blot analysis of poly A+ RNA isolated from solid tumors revealed a tetranectin specific mRNA band. In situ hybridizations on tissue sections of colon carcinomas and normal colon tissues revealed a strong and distinct hybridization signal of stromal cells in colon carcinomas but not in tumor cells. Only a few stromal cells were labeled in the normal colon. Immunohistochemically, tetranectin was found in a fibrillar-like pattern in the extracellular matrix around the tumor islands and was not detectable in the normal colon stromal tissue. Plasminogen exhibited a similar immunohistochemical staining pattern as tetranectin.

Human tetranectin cDNA comprises 874 base pairs including a 606-base pair open reading frame encoding 202 amino acids including a classical signal peptide. This protein is produced locally by cells of the stromal compartment of tumors and is deposited into the extracellular matrix. Since tetranectin binds to plasminogen we hypothesize that it could function as an anchor and/or reservoir for plasminogen and similar substances that regulate tumor invasion and metastasis as well as tumor angiogenesis.