Purification and structure of a novel cysteine proteinase inhibitor, strepin P-1.

Abstract

Strepin P-l, a new proteinase inhibitor, is a low molecular weight peptide isolated from the culture fluid of Streptomyces tanabeensis (SAB-934). Strepin P-l strongly inhibited not only cysteine proteinases, calpain and papain, but also trypsin. The purification procedures included HP-20 adsorption chromatography, DEAE-cellulose, Amberlite CG-50, Sephadex LH-20 and G-25 column chromatography. The yield was 12 mg from 8 liters of culture fluid. The proteinase inhibitor thus prepared was a peptide composed of tyrosine, valine and argininal, that reacted positively with Sakaguchi and Pauly reagents on TLC. The N-terminal amino acid, tyrosine, was blocked with an isovaleryl group and the structure was elucidated to be AMsovaleryl-tyrosyl-valyl-argininal. The amino acid sequence-inhibitory activity relationships of strepin P-l , leupeptin and antipain toward calpain and papain are also discussed.

A Survey of Calpain Inhibitors

“…Peptidyl aldehyde, diketopiperazine, and pyrazinone derivatives are among the calpain inhibitors isolated from Streptomyces species. Leupeptin (Ac-Leu-Leu-Arg-H) [30], antipain ([(S)-1-carboxy-2-phenylethyl]-carbamoyl-L-ValArg-H) [31], strepin P-1 (N-i-valeryl-Tyr-Val-Arg-H) [32] and the pentapeptides staccopins P-1 and P-2 (Val-Val-Val-Val-Phe-H and Val-Val-ValVal-Tyr-H, respectively) [33] are examples of peptidyl aldehyde calpain inhibitors isolated from Streptomyces. The inhibitors inactivate calpain by reacting reversibly with the active-site thiol of the enzyme but they are not selective for calpain.…”

A Novel Proteinase Inhibitor, Tyrostatin, Inhibiting Some Pepstatin-insensitive Carboxyl Proteinases

“…Some of them are leupeptin,39) chymostatin,40) antipain,41) MAPI,42) E_64,43) thiolstatin,44) strepin Pl, 45) and Staccopin PI and P2. 46 ) They are all peptides having special structures such as Cterminal argininal, phenylalaninal, or tyrosinal.…”

Purification and Structure of Novel Cysteine Proteinase Inhibitors, Staccopins P1 and P2, from Staphylococcus tanabeensis

“…It may catalyze limited proteolysis of a target protein or proteins in some cells, causing significant changes in metabolic processesy, 12) In an attempt to isolate new inhibitors of calpain from microbes, we isolated strepin P-l from a new strain of Streptomycetes. 6 ) t To whom all correspondence should be addressed. Cysteine proteinase inhibitors isolated by screening a number of microbes include E-64 from Aspergillus japonicus (TPR-64)13) and thiolstatin from Bacillus cereus.…”