Inhibition of human μ-calpain by conformationally constrained calpastatin peptides

Pfizer, Jose; Assfalg-Machleidt, Irmgard; Machleidt, Werner and Schaschke, Norbert (2008): Inhibition of human μ-calpain by conformationally constrained calpastatin peptides. In: Biological Chemistry, Vol. 389, No. 1: pp. 83-90

Abstract

The 27-mer peptide CP1 B-{[}1-27] derived from exon 1B of calpastatin stands out among the known inhibitors for μ- and m-calpain due to its high potency and selectivity. By systematical truncation, a 20-mer peptide, CP1B-{[}4-23], was identified as the core sequence required to maintain the affinity/selectivity profile of CP1B-{[}1-27]. Starting with this peptide, the turn-like region Glu(10)(i)-Leu(11)(i+1)-Gly(12)(i+2)-Lys(13)(i +3) was investigated. Sequence alignment of subdomains 1B, 2B, 3B and 4B from different mammalians revealed that the amino acid residues in position i+1 and i+2 are almost invariably flanked by oppositely charged residues, pointing towards a turn-like conformation stabilized by salt bridge/H-bond interaction. Accordingly, using different combinations of acidic and basic residues in position i and i+3, a series of conformationally constrained variants of CP1 B-{[}4-23] were synthesized by macrolactamization utilizing the side chain functionalities of these residues. With the combination of Glu(i)/Dab(i+3), the maximum of conformational rigidity without substantial loss in affinity/selectivity was reached. These results clearly demonstrate that the linear peptide chain corresponding to subdomain 1B reverses its direction in the region Glu(10)-Lys(13) upon binding to μ-calpain, and thereby adopts a loop-like rather than a tight turn conformation at this site.

• Item Type: Journal article
• Form of publication: Publisher's Version
• Faculties: Medicine
• Subjects: 500 Science > 540 Chemistry
• ISSN: 1431-6730
• Alliance/National Licence: This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.
• Language: English