Amino acid sequence of a trypsin inhibitor from a Spirometra (Spirometra erinaceieuropaei).
A trypsin inhibitor that is highly homologous with bovine pancreatic trypsin inhibitor (BPTI) was co-purified along with RNase from Spirometra (Spirometra erinaceieuropaei). The amino acid sequence of this inhibitor (SETI) and the nucleotide sequence of the cDNA encoding this protein were determined by protein chemistry and gene technology. SETI contains 68 amino acid residues and has a molecular mass of 7,798 Da. SETI has 31 amino acid residues that are identical with BPTI's sequence, including 6 half-cystine and 5 aromatic amino acid residues. The active site Lys residue in BPTI is replaced by an Arg residue in SETI. SETI is an effective inhibitor of trypsin and moderately inhibits a-chymotrypsin, but less inhibits elastase or subtilisin. SETI was expressed by E. coli containing a PelB vector carrying the SETI encoding cDNA; an expression yield of 0.68 mg/l was obtained. The phylogenetic relationship of SETI and the other BPTI-like trypsin inhibitors was analyzed using most likelihood inference methods.
Sanda
A
A
College of Environmental Science, Azabu University, Sagamihara, Kanagawa, Japan. sanda@azabu-u.ac.jp
Uchida
A
A
Itagaki
T
T
Kobayashi
H
H
Inokuchi
N
N
Koyama
T
T
Iwama
M
M
Ohgi
K
K
Irie
M
M
D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000737:chemistry; D001483:Base Sequence; D002417:Cattle; D018076:DNA, Complementary / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013146:Spirometra / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
D018076:DNA, Complementary; D015801:Helminth Proteins; D014361:Trypsin Inhibitors; D007611:Aprotinin