Synthesis of peptides of arginine chloromethyl ketone. Selective inactivation of human plasma kallikrein
Synthetic procedures have been developed for the preparation of peptides of arginine chloromethyl ketone and applied in the preparation of affinity labels which correspond to the -Pro-Phe-Arg- C terminus of bradykinin, a physiological cleavage site of kallikrein in kininogen. Two such reagents, Ala-Phe-ArgCH/sub 2/Cl and Pro-Phe-ArgCH/sub 2/Cl, proved to be highly effective as well as selective affinity labels for human plasma kallikrein. For example, Pro-Phe-ArgCH/sub 2/Cl inactivates plasma kallikein 50% in 24 min at a concentration of 2 x 10/sup -8/ M, while other trypsin-like proteases are less susceptible in inactivation than kallikrein, differing by a factor of 48 for plasmin and factors of 10/sup 2/ to 10/sup 5/ for factor X/sub a/, thrombin, and urokinase. The affinity of human plasma kallikrein for Ala-Phe-ArgCH/sub 2/Cl (K/sub i/ = 0.078 ..mu..M) is about 60 times that for Ala-Phe-LysCH/sub 2/Cl (K/sub i/ = 4.9 ..mu..M), whereas human plasmin exhibits about the same affinity for the former affinity label (K/sub i/ = 1.3 ..mu..M) as for the latter (K/sub i/ = 0.83 ..mu..M). The rate constants for the irreversible step of the affinity labeling reaction, k/sub 2/, are similar for all affinity labels tested with the individual proteases: 0.35 min/sup -1/ for plasma kallikrein and 0.18 min/sup -1/ for plasmin.
- Research Organization:
- Brookhaven National Lab., Upton, NY
- OSTI ID:
- 5828301
- Journal Information:
- Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 17:22; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
551000 -- Physiological Systems
59 BASIC BIOLOGICAL SCIENCES
ALANINES
AMINES
AMINO ACIDS
ANIMALS
ARGININE
AZOLES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD COAGULATION FACTORS
BLOOD PLASMA
BODY FLUIDS
BRADYKININ
CARBOXYLIC ACIDS
DRUGS
ENZYME INHIBITORS
ENZYMES
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
HYDROLASES
INACTIVATION
KALLIKREIN
KETONES
KINETICS
KININS
MAMMALS
MAN
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PEPTIDE HYDROLASES
PEPTIDES
PHENYLALANINE
PRIMATES
PROLINE
PROTEINS
PYRROLES
PYRROLIDINES
RADIOPROTECTIVE SUBSTANCES
REACTION KINETICS
RESPONSE MODIFYING FACTORS
SYNTHESIS
THROMBIN
TRYPSIN
UROKINASE
VERTEBRATES