Rabbit polymorphonuclear neutrophils form 35S-labeled S-sulfo-calgranulin C when incubated with inorganic [35S]sulfate.
Rabbit peritoneal polymorphonuclear neutrophils reduced inorganic [35S]sulfate to [35S]sulfite in vitro, concomitant with incorporation of 35S into a 10.68-kDa cytosolic protein as a S-[35S]sulfo-derivative. Amino-terminal sequencing of the purified protein identified calgranulin C, a member of the S100 protein family. cDNA clones of calgranulins B and C were isolated using oligonucleotide primers based on the established amino acid sequences of other mammalian calgranulins. The complete amino acid sequence of rabbit calgranulin C was deduced from the nucleotide sequence of the corresponding cDNA. It comprises 91 amino acid residues, has a calculated molecular mass of 10.52 kDa, has 74% identity with porcine calgranulin C, and shows high homology with other S100 calcium-binding proteins. Rabbit calgranulin C has a single cysteine residue at position 30, which we believe to be modified to S-[35S]sulfo-cysteine as a consequence of sulfate reduction by neutrophils. The formation of S-[35S]sulfo-calgranulin C appears to be a reaction specific to neutrophils. The specific radioactivity of calgranulin C from the neutrophil culture medium was 50-fold greater than that of the calgranulin C within the cells, suggesting that S-sulfation of calgranulin C might be associated with its secretion.
Yang
Z
Z
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3168, Australia.
deVeer
M J
MJ
Gardiner
E E
EE
Devenish
R J
RJ
Handley
C J
CJ
Underwood
J R
JR
Robinson
H C
HC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D040502:Calgranulin B; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D009418:S100 Proteins; D000067616:S100A12 Protein; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013431:Sulfates / Q000378:metabolism*; D013447:Sulfites / Q000378:metabolism
D002135:Calcium-Binding Proteins; D040502:Calgranulin B; D017931:DNA Primers; D009418:S100 Proteins; D000067616:S100A12 Protein; C090441:S100A12 protein, human; D013431:Sulfates; D013447:Sulfites; D003545:Cysteine