The kinetics of hydrolysis of some extended <i>N</i>-aminoacyl-<scp>l</scp>-arginine methyl esters by porcine pancreatic kallikrein. A comparison with human plasma Kallikrein

Levison, Peter R.; Tomalin, Geoffrey

doi:10.1042/bj2030299

Cited by 7 publications

(8 citation statements)

References 13 publications

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“…No indication of substrate activation which might explain our higher values for the kinetic constants for Ac-Gly-ArgOMe in comparison to recently reported data [38] was seen in our experiments. Essentially the same values were obtained when only initial velocities determined at substrate concentrations not higher than 1 mM in the case of Ac-Gly-Arg-OMe (and 0.5 mM in the case of Z-Pro-Gly-Arg-OMe) were used for the calculations (data not shown).…”

Section: Effects Of a P2 Phenylalaninelglycine Exchange In Peptide Ancontrasting

confidence: 55%

“…Morihara and Oka [72] pointed out that large effects of secondary binding are due to poor binding of a PI residue in the specificity pocket of serine proteinases. The much lower values of k,,,/K, and higher values of K, for the hydrolysis of Ac-Gly-Arg-OMe and still more Ac-Gly-Arg-Ser-Val-Gln by tissue kallikrein in comparison to those for tryptic hydrolysis, as well as the unfavourable kinetic constants for Ac-ArgOMe [38], might be interpreted as reflecting the comparatively low affinity of the S1 subsite of tissue kallikrein for arginine, due to the difference in architecture of this site 1111. However, according to Chen and Bode [ll], a small P2 residue could create an energetically unfavourable empty hydrophobic cavity at the S 2 subsite of porcine tissue kallikrein.…”

Section: Moleculur Basis Of the Secondary S2 Specificity Of Tissue Kamentioning

confidence: 99%

“…Our preliminary kinetic data for dipeptide esters with phenylalanine or glycine in P2 [12] were recently redetermined by Levison and Tomalin [38], who extended their studies also to tripeptide ester substrates. While their results for compounds with phenylalanine in P2 appear comparable to ours with due regard to the differences in pH and temperature of the measurements, the uncommented conspicuous difference expecially in k,,,, but also in K,, for Ac-Gly-Arg-OMe could not be traced to flaws in our experiments.…”

Section: Effects Of P 2 / S 2 Interactions In Tissue Kallikreinmentioning

confidence: 99%

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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: Effects Of a P2 Phenylalaninelglycine Exchange In Peptide Ancontrasting

confidence: 55%

Section: Moleculur Basis Of the Secondary S2 Specificity Of Tissue Kamentioning

confidence: 99%

Section: Effects Of P 2 / S 2 Interactions In Tissue Kallikreinmentioning

confidence: 99%

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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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“…These extraordinary features have led to studies of the substrate specificity of porcine pancreatic kallikrein with a number of synthetic peptide substrates. The majority of the substrates tested have been ethyl and methyl esters of arginine or lysine and peptides containing these residues (Levison & Tomalin, 1982b). Chloromethyl ketone inhibitors designed as either natural or synthetic substrate analogues have been succes-fully investigated as well (Fiedler et al, 1977;Kettner & Shaw, 1978).…”

mentioning

confidence: 99%

Peptide thioesters and 4-nitroanilides as substrates for porcine pancreatic kallikrein

Powers¹,

McRae²,

Tanaka³

et al. 1984

Biochemical Journal

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A series of 14 4-nitroanilide substrates and 17 thioester substrates have been used to measure kinetic constants with porcine pancreatic kallikrein. All of the substrates have a P1 arginine residue. The 4-nitroanilide substrates consist of seven P2-glycine and seven P2-phenylalanine tripeptides. As expected from previous results, the phenylalanine series substrates were generally 100-fold 'better' than those in the glycine series. The S3 subsite was found to 'prefer' lysine or phenylalanine, whereas glutamic acid in this position was distinctly unfavourable. The thioester substrates consisted of various thioester derivatives of arginine as well as 12 dipeptides. These substrates exhibited kcat./Km values generally 1000 times higher than the P2-phenylalanine 4-nitroanilides. With the thioesters, a P2 phenylalanine or tryptophan residue yielded the best substrates, but some of the simple derivatives of arginine were nearly as good. A comparison of the kinetic constants of the thioester substrates between the porcine enzyme and human plasma kallikrein provides further evidence that these enzymes have a similar preference for bulky P2 residues, but otherwise are quite different enzymes. The thioester substrates are nearly as reactive as oxygen ester substrates such as acetylphenylalanylarginine methyl ester for the porcine enzyme [Levison & Tomalin (1982) Biochem. J. 203, 299-302; Fiedler (1983) Adv. Exp. Med. Biol. 156A, 263-274], and owing to the greater ease in assaying with the thioesters, they should find use in routine assays for the glandular kallikreins.

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“…The highest activity measured was calculated to be 15 CTA-U/mg. Two active forms of plasma kallikrein have been reported (Mandle & Kaplan 1977;Nagase & Barrett 1981;Levison & Tomalin 1982a;van der Graaf et al 1982), namely kallikrein I (Mr 88,000) and kallikrein I1 (Mr 85,000). A recent report (Levison & Tomalin 1982b) on differences in the catalytic activities of the two kallikreins might support the assumption that the abovementioned differences in PGA activities could be due to the presence in the preparations of different relative amounts of the two enzymes.…”

Section: Discussionmentioning

confidence: 99%

Reduced Cofactor Function of Human High Molecular Weight Kininogen Induced by Human Plasma Kallikrein

Johansen

Briseid

1984

Acta Pharmacologica et Toxicologica

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Most reports in the literature state that human plasma kallikrein does not destroy the capacity of human high molecular weight kininogen (HMrK) to function as a cofactor in the contact phase activation of factor XII. In the present work preparations of highly purified human plasma kallikrein that showed high plasminogen activator (PGA) activities rapidly reduced the cofactor function of human HMrK. Gel electrophoresis with SDS without reduction showed that all kallikrein preparations tested contained two protein bands, one major band with a Mr of about 83,000, and one weak band with a Mr of 80,000. The main band is probably identical with kallikrein I, which Levison & Tomalin (1982b), using Ac‐Pro‐Phe‐Arg‐OMe‐HCl as substrate, found to be ten times more active (in terms of kcat/Km) than kallikrein II with Mr 3000 daltons lower. The rate of HMrK destruction in our experiments varied with the kallikrein preparation used, but assays of their hydrolytic activities against benzoyl arginine ethylester (BAEe) or the plasma kallikrein selective tripeptide substrate H‐D‐Pro‐Phe‐Arg‐pNA (S‐2302) did not discriminate between enzyme preparations with different HMrK‐destroying capacities. Assay of PGA activities demonstrated a correlation between the level of PGA measured, and the HMrK‐destroying capacity.

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The kinetics of hydrolysis of some extended N-aminoacyl-l-arginine methyl esters by porcine pancreatic kallikrein. A comparison with human plasma Kallikrein

Cited by 7 publications

References 13 publications

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

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

Peptide thioesters and 4-nitroanilides as substrates for porcine pancreatic kallikrein

Reduced Cofactor Function of Human High Molecular Weight Kininogen Induced by Human Plasma Kallikrein

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