Recovery of Argininosuccinate Lyase Activity in Duck δ1 Crystallin†

Biochemistry ◽  
2005 ◽  
Vol 44 (25) ◽  
pp. 9034-9044 ◽  
Author(s):  
May Tsai ◽  
Jason Koo ◽  
P. Lynne Howell
Keyword(s):  
Argininosuccinate Lyase ◽  
Lyase Activity

scholarly journals Inactivation of the endogenous argininosuccinate lyase activity of duck δ-crystallin by modification of an essential histidine residue with diethyl pyrocarbonate

1993 ◽  
Vol 293 (2) ◽  
pp. 537-544 ◽  
Author(s):  
H J Lee ◽  
S H Chiou ◽  
G G Chang
Keyword(s):  
Enzyme Activity ◽  
Rate Constant ◽  
Order Rate Constant ◽  
Histidine Residue ◽  
Argininosuccinate Lyase ◽  
First Order ◽  
Diethyl Pyrocarbonate ◽  
Order Rate ◽  
Lyase Activity ◽  
Pseudo First Order

The argininosuccinate lyase activity of duck delta-crystallin was inactivated by diethyl pyrocarbonate at 0 degrees C and pH 7.5. The inactivation followed pseudo-first-order kinetics after appropriate correction for the decomposition of the reagent during the modification period. The plot of the observed pseudo-first-order rate constant versus diethyl pyrocarbonate concentration in the range of 0.17-1.7 mM was linear and went through the origin with a second-order rate constant of 1.45 +/- 0.1 M-1.s-1. The double-logarithmic plot was also linear, with slope of 1.13, which suggested a 1:1 stoichiometry for the reaction between diethyl pyrocarbonate and delta-crystallin. L-Arginine, L-norvaline or L-citrulline protected the argininosuccinate lyase activity of delta-crystallin from diethyl pyrocarbonate inactivation. The dissociation constants for the delta-crystallin-L-arginine and delta-crystallin-L-citrulline binary complexes, determined by the protection experiments, were 4.2 +/- 0.2 and 0.12 +/- 0.04 mM respectively. Fumarate alone had no protective effect. However, fumarate plus L-arginine gave synergistic protection with a ligand binding interacting factor of 0.12 +/- 0.02. The double-protection data conformed to a random Uni Bi kinetic mechanism. Fluorescence-quenching studies indicated that the modified delta-crystallin had minimum, if any, conformational changes as compared with the native delta-crystallin. Inactivation of the enzyme activity was accompanied by an increasing absorbance at 240 nm of the protein. The absorption near 280 nm did not change. Treatment of the modified protein with hydroxylamine regenerated the enzyme activity to the original level. These results strongly indicated the modification of an essential histidine residue. Calculation from the 240 nm absorption changes indicated that only one histidine residue per subunit was modified by the reagent. This super-active histidine residue has a pKa value of approximately 6.8 and acts as a general acid-base catalyst in the enzyme reaction mechanism. Our experimental data are compatible with an E1cB mechanism [Raushel (1984) Arch. Biochem. Biophys. 232, 520-525] for the argininosuccinate lyase with the essential histidine residue close to the arginine-binding domain of delta-crystallin. L-Citrulline, after binding to this domain, might form an extra hydrogen bond with the essential histidine residue.

scholarly journals Staining of argininosuccinate lyase activity in polyacrylamide gel

1992 ◽  
Vol 24 (3-4) ◽  
pp. 205-214 ◽  
Author(s):  
Hwei-Jen Lee ◽  
Shyh-Horng Chiou ◽  
Gu-Gang Chang
Keyword(s):  
Polyacrylamide Gel ◽  
Argininosuccinate Lyase ◽  
Lyase Activity

Interference with the citrulline-based nitric oxide synthase assay by argininosuccinate lyase activity inArabidopsisextracts

FEBS Journal ◽  
2007 ◽  
Vol 274 (16) ◽  
pp. 4238-4245 ◽  
Author(s):  
Rudolf Tischner ◽  
Mary Galli ◽  
Yair M. Heimer ◽  
Sarah Bielefeld ◽  
Mamoru Okamoto ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Argininosuccinate Lyase ◽  
Lyase Activity ◽  
Oxide Synthase

scholarly journals Recovery of argininosuccinate lyase activity in duck δ1 crystallin

2005 ◽  
Vol 61 (a1) ◽  
pp. c186-c186
Author(s):  
M. Y. W. Tsai ◽  
J. Koo ◽  
L. M. Sampaleanu ◽  
C. Greene ◽  
L. Creagh ◽  
...  
Keyword(s):  
Argininosuccinate Lyase ◽  
Lyase Activity

scholarly journals Adaptive enzyme changes in liver and muscle of rats during protein depletion and refeeding

1968 ◽  
Vol 22 (2) ◽  
pp. 153-163 ◽  
Author(s):  
Joan M. L. Stephen
Keyword(s):  
Amino Acid ◽  
Female Rats ◽  
Male Rats ◽  
Casein Diet ◽  
Stock Diet ◽  
Argininosuccinate Lyase ◽  
Depletion Period ◽  
Lyase Activity ◽  
Diet Control ◽  
Enzyme Changes

1. Young rats were kept on a 6% casein diet for 4–7 weeks and then either killed or returned to a stock diet. Control rats were given a stock diet.2. Argininosuccinate lyase (EC 4.3.2.1, L-argininosuccinate arginine-lyase) was assayed in liver, and amino acid activating enzymes were assayed in liver and muscle of rats after depletion and while they were refed on the stock diet. Levels of enzymes were compared with those in the control rats.3. Amino acid activating enzymes were markedly increased in liver at the end of the depletion period, but returned almost to normal levels after 4–6 days of refeeding. In muscle the enzyme levels were not decreased after depletion, but began to rise after 4 days of refeeding.4. Argininosuccinate lyase activity in depleted female rats was about half that in control rats and rose again slowly on refeeding. Male rats were hardly affected by depletion.5. It is suggested that these enzyme changes play a part in the adaptation of the rat to a reduced protein intake.

scholarly journals Reversed argininosuccinate lyase activity in fumarate hydratase-deficient cancer cells

2013 ◽  
Vol 1 (1) ◽  
pp. 12 ◽  
Author(s):  
Liang Zheng ◽  
Elaine D MacKenzie ◽  
Saadia A Karim ◽  
Ann Hedley ◽  
Karen Blyth ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Fumarate Hydratase ◽  
Argininosuccinate Lyase ◽  
Lyase Activity

scholarly journals Chemical mechanism of the endogenous argininosuccinate lyase activity of duck lens δ2-crystallin

1998 ◽  
Vol 333 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Chi-Yue WU ◽  
Hwei-Jen LEE ◽  
Shih-Hsiung WU ◽  
Shui-Tein CHEN ◽  
Shyh-Horng CHIOU ◽  
...  
Keyword(s):  
Bond Cleavage ◽  
Isotope Effects ◽  
Enzymic Activity ◽  
Chemical Mechanism ◽  
Binary Complex ◽  
Argininosuccinate Lyase ◽  
Pka Values ◽  
Diethyl Pyrocarbonate ◽  
Lyase Activity ◽  
Ph Range

The endogenous argininosuccinate lyase activity of duck δ2-crystallin was specifically inactivated by the histidine-specific reagent, diethyl pyrocarbonate. The protein was protected by l-citrulline or l-arginine from the diethyl pyrocarbonate inactivation. To characterize further the chemical mechanism of the δ2-crystallin-catalysed reaction, deuterium-labelled argininosuccinate was enzymically synthesized from fumarate and l-arginine with δ2-crystallin in 2H2O. The argininosuccinate synthesized contained about 19% of the anhydride form; however, the deuterium was clearly demonstrated to be incorporated enantioselectively. Only the pro-HR atom at C-9 of the succinate moiety was labelled in the [2H]argininosuccinate-9-d synthesized, which indicates an anti-elimination mechanism for the endogenous argininosuccinate lyase activity of δ2-crystallin. The enzymic activity of duck lens δ2-crystallin in the pH range 5.5–8.5 was investigated using both protium- and deuterium-labelled argininosuccinate as the substrate. From the log kcat versus pH plot, two molecular pKa values of 6.18±0.02 and 8.75±0.03 were detected in the δ2-crystallin–argininosuccinate binary complex. The former must be dehydronated and the latter hydronated to achieve an optimum reaction rate. The log kcat/Km versus pH plot suggested two molecular pKa values of 5.96±0.09 and 8.29±0.10 for the free δ2-crystallin to be involved in the substrate binding. Small kinetic isotope effects of 1.17±0.02 and 1.05±0.09 were found for kcat and kcat/Km respectively. Combining results from labelling and kinetic analysis indicates that the endogenous argininosuccinate lyase activity of duck δ2-crystallin is compatible with a stepwise E1cB mechanism, the rate-limiting step probably at the C–N bond-cleavage step.

A Duck δ1 Crystallin Double Loop Mutant Provides Insight into Residues Important for Argininosuccinate Lyase Activity†,‡

Biochemistry ◽  
2004 ◽  
Vol 43 (37) ◽  
pp. 11672-11682 ◽  
Author(s):  
May Tsai ◽  
Liliana M. Sampaleanu ◽  
Caroline Greene ◽  
Louise Creagh ◽  
Charles Haynes ◽  
...  
Keyword(s):  
Double Loop ◽  
Argininosuccinate Lyase ◽  
Lyase Activity ◽  
Insight Into
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