Comparative studies on the primary structure of human cystatin as from epidermis, liver, spleen, and leukocytes.
We have studied the primary structure of human cystatin As from epidermis, liver, spleen, and leukocytes. These molecules were indistinguishable on PAGE in the presence and absence of SDS, by fast protein liquid chromatography (FPLC) chromatofocusing on a Mono P column, and in amino acid composition. The NH2- and COOH-terminal amino acid sequences of human cystatin As from epidermis, liver, and spleen were identical with those of human leukocyte cystatin A previously reported except for the lack of the NH2-terminal methionine residue in human epidermal cystatin A. The peptides obtained upon digestion of four human cystatin As with Achromobacter protease I (AP) showed identical peptide maps on HPLC except for different retention times of the NH2-terminal peptides. Furthermore, the amino acid compositions of corresponding separated peptide quartets were identical. We also determined the complete amino acid sequence of human epidermal cystatin A by sequencing peptides obtained from AP digestion and cyanogen bromide (CNBr) cleavage. It consisted of 97 amino acid residues, and was identical with those of human cystatin As from liver, spleen, and leukocytes except for the lack of the NH2-terminal methionine residue.
Takeda
A
A
Department of Clinical Pathology, Showa University Fujigaoka Hospital, Showa University School of Medicine, Kanagawa.
Kaji
H
H
Nakaya
K
K
Nakamura
Y
Y
Samejima
T
T
D000594:Amino Acid Oxidoreductases / Q000032:analysis* / Q000037:antagonists & inhibitors; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D001798:Blood Proteins; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D055312:Cystatin A; D015891:Cystatins; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007962:Leukocytes / Q000201:enzymology*; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000032:analysis* / Q000097:blood; D011506:Proteins / Q000032:analysis*; D012867:Skin / Q000201:enzymology*; D013154:Spleen / Q000201:enzymology*
D000596:Amino Acids; D001798:Blood Proteins; D055312:Cystatin A; D015891:Cystatins; D011480:Protease Inhibitors; D011506:Proteins; C026407:CSTA protein, human; D000594:Amino Acid Oxidoreductases; C034168:lysine epsilon-dehydrogenase; D003488:Cyanogen Bromide