Isolation of a new member of the S100 protein family: amino acid sequence, tissue, and subcellular distribution.
A low molecular mass protein which we term S100L was isolated from bovine lung. S100L possesses many of the properties of brain S100 such as self association, Ca++-binding (2 sites per subunit) with moderate affinity, and exposure of a hydrophobic site upon Ca++-saturation. Antibodies to brain S100 proteins, however, do not cross react with S100L. Tryptic peptides derived from S100L were sequenced revealing similarity to other members of the S100 family. Oligonucleotide probes based on these sequences were used to screen a cDNA library derived from a bovine kidney cell line (MDBK). A 562-nucleotide cDNA was sequenced and found to contain the complete coding region of S100L. The predicted amino acid sequence displays striking similarity, yet is clearly distinct from other members of the S100 protein family. Polyclonal and monoclonal antibodies were raised against S100L and used to determine the tissue and subcellular distribution of this molecule. The S100L protein is expressed at high levels in bovine kidney and lung tissue, low levels in brain and intestine, with intermediate levels in muscle. The MDBK cell line was found to contain both S100L and the calpactin light chain, another member of this protein family. S100L was not found associated with a higher molecular mass subunit in MDBK cells while the calpactin light chain was tightly bound to the calpactin heavy chain. Double label immunofluorescence microscopy confirmed the observation that the calpactin light chain and S100L have a different distribution in these cells.
Glenney
J R
JR
Molecular Biology and Virology Laboratory, Salk Institute for Biological Studies, San Diego, California 92138.
Kindy
M S
MS
Zokas
L
L
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins; D001483:Base Sequence; D001923:Brain Chemistry; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000032:analysis; D002417:Cattle; D002460:Cell Line; D002630:Chemotactic Factors; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D008168:Lung / Q000378:metabolism*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009132:Muscles / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D009418:S100 Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014018:Tissue Distribution
D017302:Annexins; D002135:Calcium-Binding Proteins; D002630:Chemotactic Factors; D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D009418:S100 Proteins; C098997:S100A2 protein, human; D004247:DNA; D002118:Calcium