Effect of Ca2+ on the secondary structure of linear and cyclic collagen sequence analogs.
To investigate the role of secondary structure in the substrate specificity of human 72 kDa type IV collagenase, we synthesised linear and cyclic collagen sequence analogs. As Ca2+ plays a crucial role in the enzyme activity, the CD and FTIR spectra of the peptides were also measured in the presence of Ca2+. Most of the linear, but none of the cyclic peptides form stable 1:1 Ca2+ complexes. The cyclic hexapeptides adopt significantly different backbone conformations comprising not only beta-turns but also the less frequent gamma-turns. Consequently, in the cyclopeptides the scissile Gly-Ile(Leu) bond is embedded into a different conformational environment, but in spite of that none of them is a substrate or an inhibitor of the enzyme. The best substrate Ac-Pro-Leu-Gly-Leu-Ala-Gly-D-Lys-OH binds Ca2+, but does not form a stable 1:1 Ca2+ complex, which suggests that instead of a folded structure an extended flexible conformation is preferred by the enzyme.
Likó
Z
Z
Research Group for Peptide Chemistry, Hungarian Academy of Sciences, Budapest, Hungary.
Botyánszki
J
J
Bódi
J
J
Vass
E
E
Majer
Z
Z
Hollósi
M
M
Süli-Vargha
H
H
D000595:Amino Acid Sequence; D002118:Calcium / Q000494:pharmacology*; D002942:Circular Dichroism; D003094:Collagen / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary / Q000187:drug effects*; D011994:Recombinant Proteins / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D013379:Substrate Specificity
D009842:Oligopeptides; D010446:Peptide Fragments; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D002118:Calcium