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J Pande

Publications and source records attributed to J Pande.

35 records · Page 2Linked to original sources
PubMed1991

The calf gamma crystallins--a Raman spectroscopic study.

The solution structures of the four major components of bovine lens gamma-crystallin, gamma s, gamma II, gamma III and gamma IV are compared using Raman spectroscopy. The spectral region sensitive to the vibrational frequencies of aromatic and sulfur containing residues and to the backbone skeletal stretching modes (500-1000 cm-1), and that reflecting secondary structure (1,000-1,700 cm-1) are strikingly similar in all four gamma-crystallin fractions. These similarities are indicative of the dominant anti-parallel beta sheet structure common to all the gamma-crystallins. A comparison of the ratios of the Raman intensities at 850 cm-1 and 830 cm-1 (I850/I830), an empirical measure of the degree of hydrogen bonding of phenolic hydroxyl groups, suggests that the tyrosine residues in all the gamma-crystallin fractions are moderately hydrogen bonded. Distinct differences in the solution structures of the gamma-crystallins were observed in the higher energy end of the vibrational Raman spectra. The sulfhydryl stretching frequencies for the gamma-crystallins exhibit complex splitting patterns in the 2,500-2,600 cm-1 region. These patterns are due to the competing effects of hydrogen bonding and S-pi interactions with neighboring aromatic residues. All five proteins exhibit multiple, but distinct, thiol frequencies, suggesting that the microenvironments of the cysteine residues in these proteins are significantly different.

Animals↗
PubMed1991

Effects of endosulfan on the blood and organ chemistry of freshwater fish, Barbus conchonius Hamilton.

Prolonged exposure (4 weeks) to 6.72 ppb of the organochlorine insecticide endosulfan induced disturbances in the blood and organ chemistry values of a common fish, Barbus conchonius. In blood the total lipids, cholesterol, and proteins were decreased in comparison to unexposed controls, while the free fatty acids (FFA), glucose, total phosphorus, and lactate were increased. Total lipids, FFA, and proteins were augmented in liver; cholesterol, in liver and ovary; and phosphorus and glycogen, in skeletal muscles. Compared to the controls, a decrease was seen in the total lipids (skeletal muscles and ovary), glycogen (liver, brain, and heart), and cholesterol (testes). Hyperlipemia, hyperproteinemia, and hyperlactemia persisted during a recovery period of 1 week in clean water following endosulfan poisoning.

Animals↗
PubMed1991

In vivo and in vitro effects of cadmium on selected enzymes in different organs of the fish Barbus conchonius Ham. (rosy barb).

1. Enzyme modulation by cadmium in selected organs of the fish, Barbus conchonius (rosy barb), was investigated in vivo (48 hr exposure to 12.6 mg/l cadmium chloride) and in vitro (10(-6) M cadmium chloride). 2. The acetylcholinesterase (AchE) activity was depressed in the gills but stimulated in the skeletal muscles and brain in vivo. The hepatic, branchial, and renal acid phosphatase (AcP) activity decreased marginally in vivo but it was significantly increased in the gut and ovary. In vitro, except for the liver, the AcP activity was depressed in the selected organs. Collaterally, gut alkaline phosphatase (AlP) was significantly inhibited but a pronounced stimulation was noted in the kidneys and ovary in vivo. In vitro, the AlP activity was conspicuously elevated in the kidneys and gut, and moderately in the gills. 3. Cadmium inhibited the glutamate-oxaloacetate and glutamate-pyruvate transaminases (GOT and GPT) in the liver, gills and kidneys in vivo. In vitro, the GOT and GPT activities were decreased in the liver, gills and kidneys. The lactic dehydrogenase (LDH) was significantly stimulated by Cd in the heart in vivo but in vitro the metal inhibited the enzyme in the gills. 4. Enzymes in the liver, followed by those in the kidneys and gills seem to be most seriously affected by Cd poisoning in this fish.

Acetylcholinesterase↗
PubMed1991

Individual and combined toxicity of common pesticides to teleost Puntius conchonius Hamilton.

Individual and combined toxicity of three pesticides, endosulfan, phosphamidon, and aldicarb was evaluated in P. conchonius. The 48 hr LC50 was 21.36 and 446.5 ppm respectively for endosulfan and phosphamidon. When tested jointly, 48 hr LC50 for different ratios of these pesticides were 0.332 (IE:3P), 0.224 (IE:1P), and 0.178 ppm (3E:1P). The cotoxicity coefficients for these combinations were 1793, 3986, and 10009, respectively. An equitoxic mixture of endosulfan, phosphamidon, and aldicarb yielded a 48 hr LC50 of 130.5 ppm. An enhanced toxic impact is indicated when the pesticides are present together rather than as individual compounds.

Aldicarb↗
PubMed1990

Use of the fish enzyme system in monitoring water quality: effects of mercury on tissue enzymes.

1. Rosy barb (Puntius conchonius) were exposed to 181 micrograms/l mercuric chloride for 48 h and the activity of acid and alkaline phosphatases (AcP and AIP), aspartate aminotransferase (AAT), alanine aminotransferase (AIAT), lactic dehydrogenase (LDH), and acetylcholinesterase (AchE) were measured in vivo in several organs. 2. The AcP activity was inhibited in the liver, gills, kidneys, and gut but stimulated in the gonads. With the exception of kidney, the AIP activity showed an increase in all the organs examined. The AAT and AIAT were generally inhibited in different organs. An increase in LDH activity occurred in the cardiac and skeletal muscles while the AchE activity was considerably lowered in the brain, gills, and liver. 3. In vitro exposure to mercury at concentrations ranging between 10(-10) and 10(-4) M, inhibited the AIP, AAT, AIAT, LDH, and AchE activities in the tissues examined. The AcP activity was also depressed in all the tissues except in the testes, in which a marginal increase was noted. 4. The in vivo and in vitro effects of Hg were not of similar quality implying sequestration of toxic cations in the intact animals.

Acetylcholinesterase↗
PubMed1989

Raman spectroscopic evidence for a disulfide bridge in calf gamma II crystallin.

Laser Raman spectroscopy has been applied to native and dithiothreitol-treated bovine cortical gamma II crystallin to examine the state of the thiol groups and the presence of a putative disulfide bridge. The data reveal significant differences in two key spectral regions. In the thiol stretching region (2500-2600 cm-1), the dithiothreitol-reduced form shows a 25% increase in the integrated Raman signal as compared to the native form. The magnitude of this increase corresponds to the presence of 1 mol of disulfide/mol of gamma II as determined both by the Raman data and the previous biochemical analysis from this laboratory. In the disulfide stretching region (500-540 cm-1), the native form shows a line near 511 cm-1 which is absent in the reduced form. Both native and reduced forms show a triple-banded thiol signal with one or more distinct shoulders, suggesting at least three and perhaps five different environments for the cysteine residues. The difference spectrum, obtained by a 1:1 computer subtraction of the native from the reduced form, indicates that the increase in thiol signal is centered around 2572 cm-1. In every other spectral region, both native and reduced gamma II forms are closely similar. These results strongly support the biochemical data reported earlier and indicate that the reduction of the single disulfide bridge is accompanied by minimal changes in secondary structure in solution.

Animals↗
PubMed1987

Interdependence of coenzyme-induced conformational work and binding potential in yeast alcohol and porcine heart lactate dehydrogenases: a hydrogen-deuterium exchange study.

Binding of NAD coenzymes to yeast alcohol dehydrogenase (YADH) and porcine heart lactate dehydrogenase (PHLDH) was studied by hydrogen-deuterium exchange with the infrared technique. Conformational changes in the enzymes specific to the coenzymes and their fragments were observed, and the pH dependence of the exchange reaction shows that it conforms to the EX-2 scheme. In both YADH and PHLDH the magnitude of the conformational change of measured by exchange retardation is considerably larger for NAD+ than for NADH. Studies with coenzyme fragments like ADP-ribose, ADP, and AMP also highlight the lack of rigorous correlation between structural features such as charge and size and their influence on exchange behavior. Ternary complexes such as YADH-NAD+-pyrazole, PHLDH-NAD+-oxalate, and PHLDH-NADH-oxamate, which mimic the transition state, have a significantly more pronounced effect on exchange rates than the corresponding binary complexes. The outstanding feature of this study is the demonstration that in the binary enzyme-coenzyme complexes the more loosely bound NAD+ is more effective in retarding exchange than the more firmly bound NADH. These differences are attributed to the unequal structural constraints exerted by the two coenzymes upon the enzymes, which translate to unequal expenditure of transconformational work in the formation of the two complexes. The opposing variation in the free energy of binding and the transconformational work expended can be viewed as an unequal partitioning of the net free energy gain resulting from the protein-ligand interaction into a binding term and that required for conformational change.

Alcohol Dehydrogenase↗
PubMed1986

Raman, infrared, and circular dichroism spectroscopic studies on metallothionein: a predominantly "turn"-containing protein.

Raman and IR spectra of rabbit liver metallothionein 1 (MT-1) containing 7 mol of either cadmium or zinc ions reveal high-lying amide III bands between 1290 and 1330 cm-1, indicative of beta-turns. A comparison of the splitting pattern in the amide III region below 1290 cm-1 and in the amide I band between 1600 and 1700 cm-1, with the normal-mode calculations of Lagant et al. [Lagant, P., Vergoten, G., Fleury, G., & Loucheux-Lefebvre, M. (1984a) Eur. J. Biochem. 139, 137-148; Lagant, P., Vergoten, G., Fleury, G., & Loucheux-Lefebvre, M. (1984b) Eur. J. Biochem. 139, 149-154; Lagant, P., Vergoten, G., Fleury, G., & Loucheux-Lefebvre, M. (1984c) J. Raman Spectrosc. 15, 421-423] and Krimm and Bandekar [Krimm, S., & Bandekar, J. (1980) Biopolymers 19, 1-29], suggests that metal-bound (holo) MT-1 consists largely of beta-turns of type II. In contrast, the metal-free (apo) protein displays a predominantly unordered conformation. The Raman spectra of the holoproteins below 1000 cm-1 are characterized by several unusual skeletal stretching and bending modes. The spectral pattern between 760 and 800 cm-1 in conjunction with the splitting of the amide I band agrees closely with the normal-mode calculations of Lagant et al. (1984b) on model peptides and is indicative of the presence of type III beta-turns (or 3(10)-helical segments) in MT-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Interaction of lysine residues with the metal thiolate clusters in metallothionein.

Metallothioneins are unique diamagnetic metal thiolate cluster proteins. Both vertebrate and invertebrate forms contain, besides their large cysteine content (30%), up to 14% lysine plus arginine. In the amino acid sequences, the basic residues are juxtaposed to cysteine residues and have been suggested to play a role in neutralizing the excess negative charge of the metal thiolate complexes [Kojima, Y., Berger, C., Vallee, B. L., & Kägi, J. H. R. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 3413-3417]. To document such a function, we compared the susceptibility of the lysine residues in cadmium and zinc metallothioneins and in the metal-free S-carboxamidomethyl derivative toward arylation by trinitrobenzenesulfonic acid. The results show an at least 20-fold lower initial rate of reaction with the metal-containing as opposed to that with the metal-free form, indicating a protective effect of metal complex formation on the lysine residues, the degree of protection being dependent on the nature of the metal. The modification of the lysine residues by trinitrobenzenesulfonic acid produces changes in the CD spectral features of the cadmium thiolate cluster structure. The lowered chemical reactivity of the lysine residues in the metal-containing form correlates with an upward displacement of their average pKa's to 10.9 from 10.3 in the metal-free S-carboxamidomethyl derivative. We attribute these effects at least in part to hydrogen bonding of the positively charged epsilon-amino groups to the thiolate ligands of the negatively charged cadmium thiolate units.

Acetylcysteine↗
PubMed1983

Picosecond kinetic absorption and fluorescence studies of bovine rhodopsin with a fixed 11-ene.

A synthetic retinal having a fixed 11-cis geometry has been used to prepare a nonbleachable analogue of bovine rhodopsin. Marked differences in the picosecond absorption and fluorescence behavior of this analogue at room temperature, compared with that of natural rhodopsin, were observed. This not only indicates that the 11-cis to trans isomerization of the retinal moiety is the crucial primary event in the photolysis of rhodopsin, but also it establishes that this isomerization must occur on the picosecond time scale or faster.

Animals↗
PubMed1982

Acid-base equilibrium of the Schiff base in bacteriorhodopsin.

Aqueous suspensions of dark-adapted bacteriorhodopsin (bR560) in the purple membrane of Halobacterium halobium are exposed to rapid jumps to high pH. Optical and resonance Raman measurements are carried out by using flow and stationary methods. Above pH congruent to 11.5 bR560 starts to be reversibly converted to a species absorbing at 460 nm (bR460) characterized by an unprotonated Schiff base chromophore. Above pH congruent to 13.0 bleaching takes place, first reversibly and subsequently irreversibly, to a species absorbing around 365 nm (bR365). This process competes with the formation of bR460. The pKa corresponding to the equilibrium (equation in text) is determined as 13.3 +/- 0.3. The value of the corresponding association rate constant determined from the reverse jumps (from pH 12.67 to pH 10 and 9.2) is ka = (3.5 +/- 0.5) X 10(11) M-1 s-1. Thus, starting with bR at pH 12.67 the reprotonation process is diffusion controlled as observed for homogeneous acid-base equilibria. The observed rate of dissociation when jumping from pH 6.5 to 12-13 is slower than that predicted by including the equilibrium (equation in text) The results imply that the Schiff base is titratable in the dark, but its accessibility to external OH- ions is limited. The limitations in the significance of the "apparent" value of pKa = 13.3 observed for the Schiff base titration are discussed in light of possible alterations in the structure of bR resulting from the parallel titration of other protein groups. It is suggested that a light-induced pKa change of at least nine units takes place during the photocycle of light-adapted bR.

Acid-Base Equilibrium↗
PubMed1981

Resonance Raman study of the primary photochemistry of bacteriorhodopsin.

Resonance Raman multicomponent spectra of the light-adapted form of bacteriorhodopsin, bRLA568, and its first photoproduct, K628, have been obtained at liquid nitrogen temperatures. The spectra of both bRLA568 and K628 could be obtained with the known sample compositions under our irradiating conditions and computer subtraction techniques. In agreement with previous results, we find that both bRLA568 and K628 contain chromophores linked to the apoprotein by protonated Schiff bases of retinal. Neither pigment form, suspended in H2O or 2H2O, compares closely to the spectral features of all-trans and 13-cis protonated and deuterated model chromophores, respectively. The data are consistent with other results, suggesting that a chromophore isomerization takes place in the bRLA568-to-K628 phototransition. However, the exact structure of the in situ chromophore would appear not to involve simple trans-to-13-cis structures found in solution.

Apoproteins↗
PubMed1980

The arginines of cytochrome c. The reduction-binding site for 2,3-butanedione and ascorbate.

The reactions of both ferric and ferrous horse heart cytochrome c with 2,3-butanedione, pH 7.5, 0.05 M bicarbonate buffer, under a variety of solution conditions, have been examined using amino acid integrity as the primary criterion. The only observable structural alteration was the modification of both the arginyl side chains. The reaction profiles were found to be dependent upon the presence or absence of borate and ascorbate. The single-phased modification of the arginyl side chains in ferrocytochrome c in the absence of borate is rendered biphasic in the presence of borate (borate/reagent ratio of 0.04) through substantial lowering of the rate of reaction of one of the two arginyl side chains. The addition of ascorbate inhibits the reaction in a competitive manner, particularly of the arginyl side chain undergoing rapid modification in its absence. The reactivity of both arginyl side chains to reagent for the ferricytochrome was appreciably larger than for ferrocytochrome c. The addition of reagent to ferricytochrome c also reduces heme iron, which is discerned from the development of a native-like spectrum in the region 500 to 600 nm. The differences in the reactivities of the arginyl side chains to 2,3-butanedione in the two valence states of heme iron are the reflection of small, but definite, conformational differences at the two sites in the two valence states of the protein. The concurrent reduction of heme iron and the modification of the arginyl side chains localizes the reduction and the reaction site for 2,3-butanedione. The inhibition by ascorbate of the reaction of one of the two arginines also identifies the binding domain. Of the two arginines, Arg-38 is suggested to be the ascorbate-binding site.

Animals↗