Striking susceptibility of a kinin-yielding pentadeca-peptide to tryptic hydrolysis

Prado, Eline S.; Sampaio, Misako U.; Galembeck, Fernando; Paiva, A. C. M.

doi:10.1016/0006-291x(76)90966-9

Cited by 12 publications

(4 citation statements)

References 13 publications

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“…Though not being strictly rate-limiting, k2 of 118 s-calculated from k,,, for Ac-Phe-Arg-Ser-Val-Gln ( Table 2, peptide 5; assuming k,,, for Ac-Phe-Arg-OMe obtained with the same trypsin preparation, 103 s-', to be identical to k 3 and disregarding possible nonproductive binding) is also higher than k3. A further example of this kind, the pair of substrates bradykinylSer-Val-Gln-Val-Ser and bradykinyl-OMe with k,,, values of 115-250 s-' (pH 8.1, 30"C), can be found among the data of Prado et al [42]. Though usually rate-limiting acylation in hydrolysis of amide bonds catalyzed by serine proteinases is assumed, this is evidently not correct for all such substrates.…”

Section: Rate-limiting Steps In the Hydrolysis Of Peptide And Peptidementioning

confidence: 89%

Effects of secondary interactions on the kinetics of peptide and peptide ester hydrolysis by tissue kallikrein and trypsin

Fiedler¹

1987

European Journal of Biochemistry

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Kinetic constants for the hydrolysis by porcine tissue /I-kallikrein B and by bovine trypsin of a number of peptides related to the sequence of kininogen (also one containing a P2 glycine residue instead of phenylalanine) and of a series of corresponding arginyl peptide esters with various apolar P2 residues have been determined under strictly comparative conditions. k,,, and kCa,/Km values for the hydrolysis of the Arg-Ser bonds of the peptides by trypsin are conspicuously high. k,,, for the best of the peptide substrates, Ac-Phe-Arg-Ser-Val-NH2, even reaches k,,, for the corresponding methyl ester, indicating rate-limiting deacylation also in the hydrolysis of a peptide bond by this enzyme. k,,,/K,,, for the hydrolysis of the peptide esters with different nonpolar L-amino acids in P2 is remarkably constant (range 1.7), as it is for the pair of the above pentapeptides with P2 glycine or phenylalanine. k,,, for the ester substrates varies fivefold, however, being greatest for the P2 glycine compounds. Obviously, an increased potential of a P2 residue for interactions with the enzyme lowers the rate of deacylation. In contrast to results obtained with chymotrypsin and pancreatic elastase, trypsin is well able to tolerate a P3 proline residue.In the hydrolysis of peptide esters, tissue kallikrein is definitely superior to trypsin. Conversely, peptide bonds are hydrolyzed less efficiently by tissue kallikrein and the acylation reaction is rate-limiting. The influence of the length of peptide substrates is similar in both enzymes and indicates an extension of the substrate recognition site from subsite S3 to at least Sjof tissue kallikrein and the importance of a hydrogen bond between the P3 carbonyl group and Gly-216 of the enzymes. Tissue kallikrein also tolerates a P3 proline residue well.In sharp contrast to the behaviour of trypsin is the very strong influence of the nature of the P2 residue in tissue-kallikrein-catalyzed reactions. kcal/K,,, varies 75-fold in the series of the dipeptide esters with nonpolar L-amino acid residues in P2, a P2 glycine residue furnishing the worst and phenylalanine the best substrate, whereas this exchange in the pentapeptides changes k,,,/K, as much as 730-fold. This behaviour, together with the high value of kcat/Km for Ac-Phe-Arg-OMe of 3.75 x lo7 M-' s-', suggests rate-limiting binding ( k , ) in the hydrolysis of the best ester substrates. The P2 residue mainly affects K,. The effects of the nature of the P2 residue on the hydrolysis of the dipeptide esters are in accordance with the proposed sandwiching of P2 residues between Tyr-99 and Trp-215 of tissue kallikrein. The pronounced secondary specificity of subsite S2 for bulky, hydrophobic amino acids appears to be a general feature of tissue kallikreins. Remarkably, the two most favourable P2 residues, phenylalanine and leucine, also occur in this position at the two tissue kallikrein cleavage sites of the natural substrate, kininogen. A comparison of the kinetic constants of the kininogen peptides with those for kallidin re...

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Section: Rate-limiting Steps In the Hydrolysis Of Peptide And Peptidementioning

confidence: 89%

Effects of secondary interactions on the kinetics of peptide and peptide ester hydrolysis by tissue kallikrein and trypsin

Fiedler¹

1987

European Journal of Biochemistry

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“…The kinetic constants for the enzymatic hydrolysis of peptide and ester bonds in kinin derivatives and other synthetic substrates by trypsin and horse urinary kallikrein (HoUK) have been determined by chemical methods as well as by bioassay of the released kinin when the substrate contained a kinin moiety [1,2]. The values of the kinetic constants obtained by either method were in good agreement.…”

Section: Kinin Derivatives As Substrates For Trypsin and Horse Urinarmentioning

confidence: 92%

“…BK-OME [2] 0.052 + 0.014 225 + 52 4.3 x 106 Lys-BK-OME [2] 0.10 + 0.019 194 +_ 31 1.9 x 10 * BAEE [3] 0.0043 14. trypsin are presented in Table 1. These values are compared to those obtained by hydrolysis of the ester bonds in BKOMe and Lys-BK-OME [2] as well as in BAEE [3] and of the peptide bonds in BK-R and Lys-BK-R [ 1]. The released ammonia was determined by the method of CHANEY and MARBACH [4].…”

Section: Kinin Derivatives As Substrates For Trypsin and Horse Urinarmentioning

confidence: 99%

“…BAEE3), TAME4), bradykinin methyl ester (BK-OMe), !ysyl-bradykinin methyl ester (Lys-BK-OMe), bradykinylSer-Val-Gln-Val-Ser (BK-R), lysyl-bradykinyl-Ser-Val-GlnVal-Ser (Lys-BK-R) [1,2]. We now report studies on the hydrolysis of the amide bond by trypsin and HoUK of bradykinyl-amide (BK-NH 2) and lysyl-bradykinyl-amide (Lys-BK-NH2).…”

Section: Kinin Derivatives As Substrates For Trypsin and Horse Urinarmentioning

confidence: 99%

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