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R Dua

Publications and source records attributed to R Dua.

25 records · Page 2Linked to original sources
PubMed1995

A structure-function study of bovine pancreatic phospholipase A2 using polymerized mixed liposomes.

A new combinatorial approach that includes the genetic variation of protein structure and the chemical modification of phospholipid structure in polymerized mixed liposomes was used to delineate the structure-function relationships in the interfacial catalysis of bovine pancreatic phospholipase A2 (PLA2). Based on previous structural and mutational studies, several bovine PLA2 mutants were generated in which a positive charge of putatively important lysyl side chains was reversed (K10E, K53E, K56E, and K116E) or neutralized (K56Q and K116Q). Kinetic parameters of bovine wild type and mutant PLA2s determined using polymerized mixed liposomes consisting of 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphoethanolamine (or -phosphoglycerol) and 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphoglycerol showed that Lys-53 is involved specifically in the interaction with a substrate bound in the active site. Also, these results showed that Lys-10 and Lys-116 are involved in the interaction of bovine PLA2 with anionic interfaces but not in the interaction with the active site-bound substrate. In particular, Lys-116 makes more significant contribution than Lys-10 by approximately 1.0 kcal/mol to the binding to anionic interfaces. Most importantly, Lys-56 was shown to participate in the interaction with both the active site-bound substrate and anionic interfaces. These findings establish Lys-56 and Lys-116 as essential residues for the binding of bovine pancreatic PLA2 to anionic interfaces. Lastly, our structure-function analysis based on the use of polymerized mixed liposomes was further supported by equilibrium binding measurements of these proteins using 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphoglycerol polymerized liposomes and by kinetic analyses using monomeric substrates, 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine and -phosphoglycerol.

Animals↗
PubMed1994

Inhibition of human secretory class II phospholipase A2 by heparin.

By means of kinetic analyses using Triton X-100/deoxycholic acid/dilauroylglycerophosphoethanolamine (4:2:1, molar ratio) mixed micelles we examined the effects of heparin on the activity of several phospholipases A2 (PLA2). Heparin avidly bound cationic PLA2s including human secretory class II PLA2 and thereby inhibited their hydrolysis of phospholipids in the mixed micelles. Initial velocity measurements indicated that heparin behaved as a competitive inhibitor for human secretory class II PLA2 and closely related A.h. blomhoffii PLA2 and A.p. piscivorus PLA2. In particular, heparin showed the highest specificity for human secretory class II PLA2. In the absence of deoxycholic acid in mixed micelles, A.h. blomhoffii PLA2 was also strongly inhibited by heparin. The observed inhibition was not due to the interaction of heparin with the active site of PLA2 because heparin did not inhibit the hydrolysis of monomeric substrates by PLA2s. Both kinetic measurements and fluorescence measurements of PLA2-bound 8-anilino-1-naphthalene sulfonate in the presence of varying amounts of heparin showed that a heparin molecule bound about seven molecules of PLA2. When positive charges of four lysines in the amino-terminal region of A.h. blomhoffii PLA2 were neutralized by limited carbamoylation, heparin neither bound the carbamoylated A.h. blomhoffii PLA2 nor inhibited the hydrolysis of Triton X-100/dilauroylglycerophosphocholine mixed micelles by the carbamoylated A.h. blomhoffii PLA2 that retained 50% activity of native A.h. blomhoffii PLA2. Also, heparin did not inhibit the hydrolysis of mixed micelles by 7,10-bis(octanoyl)ated A.p. piscivorus PLA2 in which two lysines in the amino-terminal alpha-helix are acylated. These results indicate that the inhibition of human secretory class II PLA2 and related cationic PLA2s by heparin originates from the interaction of heparin with cationic residues in the amino-terminal region that forms a part of interfacial binding site. In addition, unique structural features of human secretory class II PLA2, together with its unique mode of interaction with heparin, suggest that this PLA2 might have an additional heparin-binding site. Although the heparin-PLA2 binding diminished as the ionic strength of reaction medium increased, the inhibition of human secretory class II PLA2 by heparin remained significant at the physiological ionic strength. An estimated value of inhibition constant (Ki) was 0.1 microM under physiological conditions, which suggests that a normal pharmaceutical dose of heparin might inhibit human secretory class II PLA2 and regulate its biological effects.

Amino Acid Sequence↗
PubMed1993

Long-term follow-up after two coronary repair of anomalous left coronary artery from the pulmonary artery.

A retrospective analysis of ten patients with anomalous left coronary artery arising from the pulmonary artery operated between 1979 and 1990 was undertaken. All presented with evidence of left ventricular dysfunction and "ischemic" mitral regurgitation. Surgical repair consisted of an aortopulmonary tunnel (Takeuchi) procedure in eight and direct left coronary artery reimplantation in two. Two patients required postoperative support with a left ventricular assist device. There were no operative or late deaths (CL 0% to 17%) for a follow-up of over 670 patient months. All patients are in New York Heart Association Class I or II, though two patients are still receiving anticongestive medications. One patient has required further surgery for pulmonary artery stenosis, and another has had a mitral valve replacement because of severe mitral regurgitation. One additional patient has moderate-to-severe residual mitral regurgitation and two have a trivial left coronary to main pulmonary artery fistula. All have a patent, nonstenotic left coronary artery and much improved left ventricular function and perfusion as assessed by echocardiography, thallium scan, gated blood pool scan, and angiography. There have been no documented arrhythmias, clinically or on Holter monitoring. The ECGs have shown resolution or improvement of the initial changes of ischemia/infarction in all patients. Chest X-rays have shown normalization of cardiothoracic ratio in eight of ten patients. Excellent early and late results can be achieved following timely surgical repair. Marked improvement in left ventricular function has been observed in patients with poor preoperative left ventricular function, even in the presence of extensive ischemia/infarction.

Child, Preschool↗
PubMed1991

Dopamine receptors in the ventral tegmental area affect motor, but not motivational or reflexive, components of copulation in male rats.

Microinjection of apomorphine into the ventral tegmental area (VTA) of male rats was previously shown to delay the onset of copulation and slow its rate, presumably by stimulating impulse-regulating autoreceptors on cell bodies of the A10 mesocorticolimbic dopamine tract. Such stimulation would be expected to slow the firing rate of these neurons and, thereby, to impair locomotion and/or motivational processes. The present experiments tested whether the delayed onset and slowed rate of copulation were related to deficits in motor performance, sexual motivation, and/or genital reflexes. In X-maze tests the speed of running to all 4 goal boxes was slowed; however, the percentage of trials on which the male chose the female's goal box was not decreased. Examination of videotaped copulation tests revealed that the male showed fewer complete copulatory behaviors (mounts, intromissions, and ejaculations), but more misdirected or incomplete copulatory attempts after apomorphine in the VTA. There were also fewer scores of active, as opposed to inactive, behaviors, and the onset and rate of copulation were slowed. The total number of female directed behaviors was not different in apomorphine tests, compared to vehicle. Finally, tests of ex copula genital reflexes revealed no significant effects of apomorphine in the VTA on erections, penile movements, or seminal emissions. These data suggest a role of the VTA in the motor aspects and/or sensorimotor integration of copulation. Sexual motivation and ex copula genital reflexes appeared to be unaffected by apomorphine in the VTA.

Animals↗