Abstract
Three molecular forms of a protein inhibitor of chymotrypsin and microbial alkaline proteases have been isolated from Hiproly high-lysine barley.
Automated Edman degradation of one of these inhibitor preparations (CI-1C) resulted in the following amino acid sequence (77 residues in total): Tyr-Pro-Glu-Pro-Thr-Glu-Gly-Ser-Ile-Gly-Ala-Ser-Gly-Ala-Lys-Thr-Ser-Trp-Pro-Glu-Val-Val-Gly-Met-Ser-Ala-Glu-Lys-Ala-Lys-Glu-Ile-Ile-Leu-Arg-Asp-Lys-Pro-Asn-Ala-Gln-Ile-Glu-Val-Ile-Pro-Val-Asp-Ala-Met-Val-Pro-Leu-Asn-Phe-Asn-Pro-Asn-Arg-Val-Phe-Val-Leu-Val (His, Lys, Ala, Thr, Thr, Val, Ala, Glx, Val, Ser, Arg) Val-Gly.
The inhibitor (CI-1) is homologous with another barley inhibitor (CI-2), with potato inhibitor I and with the elastase-cathepsin G inhibitor eglin from the leech Hirudo medicinalis (30–50% of the amino acid residues in identical positions). This established «family of cystine-independent inhibitors» also showed some sequence similarities with the cystine-free yeast proteinase B inhibitors 1 and 2. In the reactive site region homologies with the cystine-rich inhibitors of the «Kazal pancreas secretory inhibitor» and the «Streptomyces subtilisin inhibitor» families were observed.
Article PDF
Similar content being viewed by others
Abbreviations
- APNE:
-
N-acetyl-d,l-phenylalanine-2-naphtylester
- CI-1 and CI-2:
-
chymotrypsin inhibitor 1 and 2 from barley (3)
- LIE:
-
Leech inhibitor eglin (17)
- PI-I:
-
potato inhibitor I (15)
- Polybrene:
-
hexadimethrine bromide
- PTH:
-
phenylthiohydantoin
- SDS:
-
sodium dodecyl sulfate
- THEED:
-
N,N,N′,N′,-tetrakis (2-hydroxyethyl)ethylenediamine
- YIB:
-
yeast inhibitor of protease B (11)
References
Begg, G. &F. J. Morgan: A non-volatile buffer with improved performance in automated protein sequencing. FEBS Lett. 66, 243–245 (1976)
Biedermann, K., U. Montali, B. Martin, I. Svendsen &M. Ottesen: The amino acid sequence of proteinase A inhibitor 3 from baker's yeast. Carlsberg Res. Commun. 45, 225–235 (1980)
Boisen, S., C. Y. Andersen &J. Hejgaard: Inhibitors of chymotrypsin and microbial serine proteases in barley grains. Isolation, partial characterization and immunochemical relationships of multiple molecular forms. Physiol. Plant. 52, 167–176 (1981)
Hejgaard, J. &S. Boisen: High-lysine proteins in Hiproly barley breeding: Identification, nutritional significance and new screening methods. Hereditas 93, 311–320 (1980)
Hejgaard, J.: Isoelectric focusing of subtilisin inhibitors: Detection and partial characterization of cereal inhibitors of chymotrypsin and microbial proteases. Anal. Biochem. 116, 444–449 (1981)
Jonassen, I.: Characteristics of Hiproly barley I. Isolation and characterization of two water-soluble high-lysine proteins. Carlsberg Res. Commun. 45, 47–58 (1980)
Kato, I., W. J. Kohr &M. Laskowski, Jr.: Evolution of avian ovomucoids. In: Regulatory proteolytic enzymes and their inhibitors. Proc. 11th FEBS Meeting, Copenhagen 1977. S. Magnusson et al., eds., Pergamon Press, Oxford, pp. 197–206 (1978)
Kominami, E., H. Hoffschulte, L. Leuschel, K. Maier &H. Holzer: The substrate specificity of proteinase B from baker's yeast. Biochim. Biophys. Acta 661, 136–141 (1981)
Kulbe, K. D.: Micropolyamide thin-layer chromatography of phenyl-thiohydantoin amino acids (PTH) at a subnanomolar level. Anal. Biochem. 59, 564–573 (1974)
Laskowski, M. Jr., &I. Kato: Protein inhibitors of proteinases. Ann. Rev. Biochem. 49, 593–626 (1980)
Maier, K., H. Müller, R. Tesch, I. Witt &H. Holzer: Amino acid sequence of yeast proteinase B inhibitor 1. Comparison with inhibitor 2. Biochem. Biophys. Res. Commun. 91, 1390–1398 (1979)
Martin, B., I. Svendsen &M. Ottesen: Use of carboxypeptidase Y for carboxy-terminal sequence determination in proteins. Carlsberg Res. Commun. 42, 99–102 (1977)
Mikola, J. &E.-M. Suolinna: Purification and properties of an inhibitor of microbial alkaline proteinases from barley. Arch. Biochem. Biophys. 144, 566–575 (1971)
Plunkett, G. &C. A. Ryan: Reduction and carboxamidomethylation of the single disulfide bond of proteinase inhibitor I from potato tubers. Effects on stability, immunological properties, and inhibitory activities. J. Biol. Chem. 255, 2752–2755 (1980)
Richardson, M. &L. Cossins: Chymotryptic inhibitor I from potatoes: The amino acid sequences of subunits B, C, and D. FEBS Lett. 45, 11–13 (1974). (Corrigendum in FEBS Lett. 52, 161 (1975))
Richardson, M., R. D. J. Barker, R. T. McMillan &L. M. Cossins: Identification of the reactive (inhibitory) sites of chymotryptic inhibitor I from potatoes. Phytochemistry 16, 837–839 (1977)
Seemüller, U., M. Eulitz, H. Fritz &A. Strobl: Structure of the Elastase-Cathepsin G inhibitor of the leechHirudo medicinalis. Hoppe-Seyler's Z. Physiol. Chem. 361, 1841–1846 (1980)
Seemüller, U., H. Fritz &M. Eulitz: Eglin: Elastase-Cathepsin G inhibitor from leeches. Methods in Enzymology 80, 804–816 (1981)
Svendsen, I., B. Martin &I. Jonassen: Characteristics of Hiproly barley. III. Amino acid sequences of two lysine-rich proteins. Carlsberg Res. Commun. 45, 79–85 (1980)
Svendsen, I., I. Jonassen, J. Hejgaard &S. Boisen: Amino acid sequence homology between a serine protease inhibitor from barley and potato inhibitor I. Carlsberg Res. Commun. 45, 389–395 (1980)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Svendsen, I., Boisen, S. & Hejgaard, J. Amino acid sequence of serine protease inhibitor CI-1 from barley. Homology with barley inhibitor CI-2, potato inhibitor I, and leech eglin. Carlsberg Res. Commun. 47, 45–53 (1982). https://doi.org/10.1007/BF02907796
Issue Date:
DOI: https://doi.org/10.1007/BF02907796