Fibrin-stabilizing factor inhibitors. 11. Monodansylated weak aliphatic diamines

Specificity of guinea pig liver transglutaminase for amine substrates

“…From Lineweaver-Burk plots of the experimental data, uncorrected ÁTM_app values of about 3 X 10~4 and 7 X 10~5 M were calculated for dansylcadaverine and dansylthiacadaverine, respectively. If, however, when the p/fa values of 10.6 and 9.6 for the primary ammonium ions of these two compounds are used (Ljunggren et al, 1974;Perrin, 1965), Michaelis constants are expressed in relation to the effective concentrations of unprotonated amine species existing at the pH of experiments (pH 7.5; 25 °C) rather than on the basis of the total (i.e., amine plus ammonium ion) concentration of substrates, a corrected ATM_app of 3 X 10"7 M is obtained for dansylcadaverine and 7 X 10~7 M is obtained for dansylthiacadaverine. These figures provide quantitative proof that both of these amines are excellent substrates for transglutaminase and that the cadaverine derivative may actually be somewhat more specific than the thiacadaverine analogue.…”

“…Dansylcadaverine was chosen as standard and its Afi app_1 value of about 4 X 107 M'1 was taken as 100% on the affinity scale. In calculating corrected relative affinities, we made the following assumptions: (a) mono-yV-acyl-and -TV-sulfonylalkyldiamine derivatives which have a five methylene-carbon spacer between the primary amino group and the amide substituent have a pXa for the primary amino group identical with that in dansylcadaverine [compound III; pXa = 10.6 ± 0.1; Perrin (1965) and Ljunggren et al (1974)]; (b) mono-/V-acyl-3thiacadaverine derivatives (where a -CH2group in position 3 of the alkyl side chain is replaced by a -S-) have the same pAfa for the primary amino group as in dansylthiacadaverine [compound XXXVI; pXa = 9.6 ± 0.1; Perrin (1965) and…”

Kinetics of transamidating enzymes. Production of thiol in the reactions of thiol esters with fibrinoligase

“…Three of the amines, dansylcadaverine (Lorand et al. 1968), dansylthiacadaverine (Ljunggren et al, 1974), and TV-a-p-tosyl-L-lysine methyl ester, are considered to be very good substrates for the enzyme; the fourth, putrescine (Lorand et al, 1972b), is of intermediate and the fifth, n-butylamine (Lorand et al, 1968), is of low specificity. While the concentration of amines necessary for half-maximal velocity of thiocholine production varied by more than two orders of magnitude within this series, the maximally obtained rates at saturating concentrations of all amines were essentially the same (1.2-2.1 sec-1).…”

“…Found: C, 56.63; , 7.31; N, 4.95. 2-Benzyldiethylaminoethanethiol cinnamate bromide (Ljunggren et al, 1973) was a gift from Dr. Christine Ljunggren of the University of Uppsala, Sweden. Amine substrates included jV-(5-aminopentyl)-5-dimethylamino-I -naphthalenesulfonamide or dansylcadaverine (Lorand et al, 1968;Nilsson et al, 1971), A'-(5-aminopentyl)-/?-toluenesulfonamide or tosylcadaverine (Lorand et al, 1968;Stenberg et al, 1972), A'-(5-amino-3-thiapentyl)-5-dimethylamino-1 -naphthalenesulfonamide or dansylthiacadaverine (Ljunggren et al, 1974), n-butylamine hydrochloride (Eastman Kodak Co.), A'-a-p-tosyl-L-lysine methyl ester hydrochloride (Cyclo Chemical Corp.), and putrescine dihydrochloride (Sigma).…”

A Specific, Fluorescent Activity Staining Procedure Applied to Plasma and Red Blood Cells in Congenital Factor XIII Deficiency

“…It does not require the use ofany complicated equipment and can thus be set up at any routine laboratory. The fluorescent amine, monodansylthiacadaverine, is easily prepared in one step (Ljunggren et al, 1974) and exhibits a large degree of specificity (Curtis et al, 1974;Stenberg et al, 1975). As even a low factor XI11 activity alone does not necessarily result in clinical haemorrhage (Duckert, 1972), the sensitivity of the assay is crucial.…”