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Biomedical subjects

M R Chance

Publications and source records attributed to M R Chance.

At least 19 recordsLinked to original sources
PubMed1992

Extended x-ray absorption fine structure studies of a retrovirus: equine infectious anemia virus cysteine arrays are coordinated to zinc.

Zinc finger arrays have been established as a critical structural feature of proteins involved in DNA recognition. Retroviral nucleocapsid proteins, which are involved in the binding of viral RNA, contain conserved cysteine-rich arrays that have been suggested to coordinate zinc. We provide metalloprotein structural data from an intact virus preparation that validate this hypothesis. Extended x-ray absorption fine structure (EXAFS) spectroscopy of well-characterized and active preparations of equine infectious anemia virus, compared with a peptide with known coordination and in combination with available biochemical and genetic data, defines a Cys3His1 coordination environment for zinc. The average of the Zn-S distances is 2.30(1) A and that of the Zn-N distance (to histidine) is 2.01(3) A.

Absorptiometry, Photon↗
PubMed1992

Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution-state structure of the nucleocapsid protein from HIV-1.

All retroviral nucleocapsid (NC) proteins contain one or two copies of an invariant 3Cys-1His array (CCHC = C-X2-C-X4-H-X4-C; C = Cys, H = His, X = variable amino acid) that are essential for RNA genome packaging and infectivity and have been proposed to function as zinc-binding domains. Although the arrays are capable of binding zinc in vitro, the physiological relevance of zinc coordination has not been firmly established. We have obtained zinc-edge extended X-ray absorption fine structure (EXAFS) spectra for intact retroviruses in order to determine if virus-bound zinc, which is present in quantities nearly stoichiometric with the CCHC arrays (Bess, J.W., Jr., Powell, P.J., Issaq, H.J., Schumack, L.J., Grimes, M.K., Henderson, L.E., & Arthur, L.O., 1992, J. Virol. 66, 840-847), exists in a unique coordination environment. The viral EXAFS spectra obtained are remarkably similar to the spectrum of a model CCHC zinc finger peptide with known 3Cys-1His zinc coordination structure. This finding, combined with other biochemical results, indicates that the majority of the viral zinc is coordinated to the NC CCHC arrays in mature retroviruses. Based on these findings, we have extended our NMR studies of the HIV-1 NC protein and have determined its three-dimensional solution-state structure. The CCHC arrays of HIV-1 NC exist as independently folded, noninteracting domains on a flexible polypeptide chain, with conservatively substituted aromatic residues forming hydrophobic patches on the zinc finger surfaces. These residues are essential for RNA genome recognition, and fluorescence measurements indicate that at least one residue (Trp37) participates directly in binding to nucleic acids in vitro. The NC is only the third HIV-1 protein to be structurally characterized, and the combined EXAFS, structural, and nucleic acid-binding results provide a basis for the rational design of new NC-targeted antiviral agents and vaccines for the control of AIDS.

Amino Acid Sequence↗
PubMed1992

Formation of a square-planar Co(I) B12 intermediate. Implications for enzyme catalysis.

X-ray edge and extended x-ray absorption fine structure (EXAFS) techniques provide powerful tools for analysis of local molecular structure of complexes in solution. We present EXAFS results for Co(I) B12 that demonstrate a four-coordinate (distorted) square-planar configuration. Comparison of EXAFS solutions for Co(I) and Co(II) B12 (collected previously; Sagi et al. 1990. J. Am. Chem. Soc. 112:8639-8644) suggest that modulation of the Co-N bond to the axial 5,6-dimethylbenzimidazole (DMB), in the absence of changes in Co-N (equatorial) bond distances, may be a key mechanism in promoting homolytic versus heterolytic cleavage. As Co-C bond homolysis occurs, the Co-N (DMB) bond becomes stronger. However, for heterolytic cleavage to occur, earlier electrochemical studies (D. Lexa and J. M. Saveant. 1976. J. Am. Chem. Soc. 98:2652-2658) and recent studies of methylcobalamin-dependent Clostridium thermoaceticum (Ragsdale et al. 1987. J. Biol. Chem. 262:14289-14297) suggest that removal of the DMB ligand (before Co-C bond cleavage) favors formation of the four-coordinate square-planar Co(I) species while inhibiting formation of the five-coordinate Co(II) B12 complex. This paper presents the first direct evidence that formation of the Co(I) B12 intermediate must involve breaking of the Co-N (DMB) bond.

Cobamides↗
PubMed1991

Identification of structural markers for vitamin B12 and other corrinoid derivatives in solution using FTIR spectroscopy.

The identification of structural markers for B12/protein interactions is crucial to a complete understanding of vitamin B12 transport and metabolic reaction mechanisms of B12 coenzymes. Fourier transform infrared spectroscopy can provide direct measurements of changes in the side chains and corrin ring resulting from B12/protein interactions. Using FTIR spectroscopy in various solvent systems, we have identified structural markers for corrinoids in the physiological state. We assign the major band (denoted B), which occurs at ca. 1630 cm-1 in D2O and ca. 1675 cm-1 in ethanol, to the amide I C=O stretching mode of the propionamide side chains of the corrin ring. The lower frequency of band B in D2O versus ethanol is due to the greater hydrogen-bonding properties of D2O that stabilize the charged amide resonance form. Since the propionamides are known to be important in protein binding, band B is a suitable marker for monitoring the interaction of these side chains with proteins. We assign bands at ca. 1575 and 1545 cm-1 (denoted C and D) as breathing modes of the corrin ring on the basis of the bands' solvent independence and their sensitivity to changes in axial ligation. As the sigma-donating strength of the axial ligands increases, the frequencies of bands C and D decrease, possibly indicating a lengthening of the corrin conjugated system. Band A, the known cyanide stretching frequency at ca. 2130 cm-1, probes the cobalt-carbon distance in cyanocorrinoids. As the frequency of band A increases, the cobalt-carbon bond strength should decrease.

Amides↗
PubMed1990

Nanosecond transient absorption spectroscopy of coenzyme B12. Quantum yields and spectral dynamics.

Photolysis of adenosylcobalamin leads to homolytic cleavage, similar to many of the B12-dependent enzyme reactions. Therefore, we have used photolysis to study the structure and lability of the cobalt-carbon bond. The nanosecond quantum yield for adenosylcobalamin is 0.23 +/- 0.04, higher than reported previously. The acidified form of adenosylcobalamin, so called "base-off" B12, has a much lower quantum yield at 0.045 +/- 0.015, demonstrating an inverse correlation between cobalt-carbon bond strength and quantum yield. Investigation of the wavelength dependence of the quantum yield shows that there is a highly efficient transmission of energy from the corrin ring to the cobalt-carbon bond. A comparison of nanosecond transient and static spectra showed small spectral differences. Therefore, any spectral relaxation of a sterically distorted corrin ring may be detectable only at sub-nanosecond timescales. Spectral analysis also provides data on the kinetics of recombination. In the absence of enzyme, geminate rebinding must be substantial, since the rate of Co(II) and deoxyadenosyl radical recombination is near the diffusion controlled limit. Therefore, it is likely that the enzyme functions to pull the geminate partners apart, perhaps as suggested previously, through a conformational change. The importance of geminate recombination in the mechanism of homolytic cleavage is further supported by a comparison of our results with picosecond transient absorption studies.

Cobamides↗
PubMed1990

O2 and CO reactions with heme proteins: quantum yields and geminate recombination on picosecond time scales.

Picosecond time-resolved absorption spectroscopy and low-temperature studies have been undertaken in order to understand the nature of the intrinsic quantum yields and geminate recombination of carbon monoxide and oxygen to hemoglobin and myoglobin. We find that the photoproduct yields at 40 ps and long times (minutes) after photolysis at 8 K are similar; however, the yield of oxygen photoproducts is 0.4 +/- 0.1 while the yield of carbon monoxide photoproducts is 1.0 +/- 0.1 for both myoglobin and hemoglobin. Measurements in the Soret, near-infrared, and far-IR are used to quantitate the photoproduct yields. These results call into question previous cryogenic kinetic studies of O2 recombination. Significant subnanosecond geminate recombination is observed in oxyhemoglobin down to 150 K, while below 100 K this geminate recombination disappears. The lower photoproduct yields for oxyheme protein complexes can be attributed to both subnanosecond and subpicosecond recombination events which are ligand and protein dynamics dependent.

Animals↗
PubMed1990

Structural and functional significance of inhomogeneous line broadening of band III in hemoglobin and Fe-Mn hybrid hemoglobins.

Near-infrared spectra of hemoglobin and Fe-Mn hybrid hemoglobins have been obtained at cryogenic temperatures. The charge-transfer (a2u(pi)----dzy) transition at approximately 760 nm (band III) has been found to be a conformationally sensitive indicator of the heme-pocket geometry in these species. Temperature, protein tertiary and quaternary structure, chain heterogeneity, and ligand rebinding subsequent to CO photolysis all affect the line width and position of this transition. We conclude that the overall line shape of band III is derived from both subunit heterogeneity and conformational disorder within each subunit. A model is suggested that relates the observed pH dependence of the kinetic hole burning due to ligand rebinding to specific structural parameters of the proximal heme pocket that influence both the peak position and the inhomogeneous line shape of band III.

Binding Sites↗
PubMed1987

Ligand binding channels reflected in the resonance Raman spectra of cryogenically trapped species of myoglobin.

Variations in the v2 region of the Raman spectra of cryogenically trapped photoproducts of different liganded myoglobins as a function of ligand (CO, O2, and n-butyl isocyanide) and species (whale, tuna, elephant) are reported. These variations are attributed to differences in the population of "open" (ligand accessible) and "closed" (ligand inaccessible) conformations of the distal heme pocket. Based on these findings and those derived from other spectroscopies including x-ray crystallography, NMR, IR spectra, and ESR, a working model is presented which accounts for how the conformation of the distal heme pocket, the geometry of the bound ligand, the identity of the ligand, and the dynamics of the dissociated ligand are all interconnected.

Animals↗
PubMed1987

Linkage of functional and structural heterogeneity in proteins: dynamic hole burning in carboxymyoglobin.

Inhomogeneous broadening of the 760-nanometer photoproduct band of carboxymyoglobin at cryogenic temperatures has been demonstrated with a dynamic hole burning technique. Line-shape changes and frequency shifts in this spectral band are generated by ligand recombination and are shown not to be the result of structural relaxation below 60 K. The observation of dynamic hole burning exposes the relation between the structural disorder responsible for the inhomogeneous broadening and the well-known distributed ligand rebinding kinetics. The findings provide direct evidence for the functional relevance of conformational substrates in myoglobin rebinding. In addition, a general protocol for evaluating the relative contributions of structural relaxation and hole burning to the spectral changes accompanying rebinding in hemeproteins is presented.

Animals↗
PubMed1987

Myoglobin recombination at low temperature. Two phases revealed by Fourier transform infrared spectroscopy.

The low-temperature recombination of CO with myoglobin was studied using Fourier transform infrared spectroscopy. The bound state of carbon monoxide myoglobin has two distinct conformers observed at 1926 and 1945 cm-1 with an intensity ratio of 1 to 8. The recombination of these bands after complete photolysis at 10 K followed by a temperature jump shows distinct kinetics for the two bands. Although both bands apparently follow the nonexponential kinetics originally described by Frauenfelder and co-workers (Austin, R., Beeson, K., Eisenstein, L., Frauenfelder, H., and Gunsalus, I. (1975) Biochemistry 14, 5355-5373), the 1926 cm-1 band does not appear appreciably below 70 K. In fact, after 20 min of recombination at 70 K the 1945 cm-1 band is fully recovered, while no detectable amount of the 1926 cm-1 band is present. This is the first association of a spectroscopic marker of protein substructure with reaction kinetics.

Carbon Monoxide↗
PubMed1986

Movement of Fe with respect to the heme plane in the R-T transition of carp hemoglobin. An extended x-ray absorption fine structure study.

Carp Hb undergoes a well known change in kinetics over the pH range 6-9. X-ray absorption spectroscopy, in conjunction with refined data analysis procedures, shows no difference in iron-ligand distances when carp HbCO is switched from R (high affinity) to T (low affinity) states. These distances are 2.015 +/- 0.015 A for the average iron-pyrrole nitrogen distance, 2.14 +/- 0.04 A for the iron-nitrogen (of histidine) distance, and 1.89 +/- 0.05 A for the Fe-C (of CO) distance. Examination of the region from 30 to 100 eV above the threshold, called the ligand field indicator region, reveals spectral differences, which when compared to model compounds suggest that the iron and the heme are less coplanar in the T-like forms. These results are consistent with the iron being 0.1 A more out of the mean heme plane in both carp HbCO and carp Hb T states, relative to the R forms, and that the change in iron position on ligation to either T or R state is four times larger than that occurring with the quaternary switch.

Animals↗
PubMed1979

An ethological perspective on social behavior in long stay hospitalized psychiatric patients.

Ethological methods were employed to gather normative data on social behavior in long stay male inpatients in the ward environment. Most were diagnosed schizophrenic. Social bheavior was categorized into four main types: altruistic, assertive, cigarette, and verbal. Patients (N = 24) were divided into three groups on the basis of the amount of social interaction (top, middle, and bottom thirds of the interactional scale). With the use of these divisions, results indicated: a) marked variability between thirds in the amount of social behavior exhibited; b) differences in the kinds of social behavior utlized by each third--the top was predominantly verbal whereas the middle and bottom were predominantly nonverbal; c) differences in the modes of interaction between thirds--verbal behavior characterized interactions between the top and top, and top and bottom, whereas assertive behavior characterized interactions between the top and middle, middle and middle, middle and bottom, and bottom and bottom; and d) correlative relationships between various social and nonsocial parameters, the most noteworthy including the positive relationship between head up, eyes open, and high rate of social interaction, and the positive relationships between altruistic, cigarette, and verbal behavior. Discussion focused on issues of variability between patients, stability of behaviors over time, behaviors indicative of bonding and social attentiveness, profiles of behavior characteristic of each third of the interactional scale, and the ethological constructs of dominance and attention structure.

Adult↗
PubMed1979

An ethological analysis of long stay hospitalized psychiatric patients. Senders and receivers in social interaction.

Ethological methods were employed to monitor social interaction among 24 long stay male patients on a psychiatric ward. Most were diagnosed as schizophrenic. Analysis was made using four molar behavioral groupings: assertive, altruistic, cigarette, and verbal. Within each grouping social behavior was divided into two categories: sending and receiving. Patients were ranked in an interactional scale and divided into three groups on the basis of the amount of social interaction: top, middle, and bottom third. Results showed that individuals in the top third tended to be senders of social behavior, whereas individuals in the middle and bottom thirds tended to be receivers. Individuals in the middle received relatively large amounts of assertive behavior. In addition, correlative relationships indicated positive associations between send and receive verbal and send and receive cigarette. No associatiociation existed between send and receive verbal and send and receive assertive. Findings are discussed in terms of a) sending and receiving profiles characteristic of each third of the interactional scale; b) the value of dividing social behavior into components of sending and receiving; and c) the role assertive behavior plays in the social organization of hospitalized groups.

Adult↗
PubMed1977

Factors influencing nursing in Macaca fascicularis.

Nursing by two mother-infant pairs in a caged colony of Macaca fascicularis was monitored at 1-min intervals for 8 h beginning 8:30 a.m. BST, once a week for 3 months in the summer of 1973. Nursing occupied about 210 min in 8 daylight hours for the infants at 10 weeks of age, and the time spent nursing decreased at the average rate of 9.4 min per week until the infants were about 6 months old. The time spent nursing by the infants studied here resembles closely the times spent nursing by some other macaques and by baboons. In the course of a day the amount of time spent nursing varies significantly with a diurnal peak. If nursing by one mother-infant pair is independent of nursing by the other pair, then the time the two pairs spend nursing together would be a function of the product of the frequencies of nursing by each pair. The expected times for the pairs nursing together based on the hypothesis of independent events were significantly less than the observed times the pairs nursed together. Nursing, therefore, involves a positive influence or imitation of one nursing pair by the other. Nursing sessions involving both mother-infant pairs were longer on the average than sessions involving only one pair.

Age Factors↗
PubMed1975

Behavioural changes in laboratory mice during cannabis feeding and withdrawal.

The effects of feeding cannabis at a level of 0.4% in the diet has been studied by an ethological analysis of encounters between male mice. Administration of cannabis to dominant males resulted in a reduction of non-social activity and an increase in flight and in social and sexual investigation when compared with untreated controls, but the behaviour of subordinate males was not significantly altered by cannabis. One week after withdrawal of cannabis, the behaviour of diminant males showed a rebound effect with increase in aggression. Nevertheless, by a preference feeding test it was demonstrated that the treated mice were not dependent on the cannabis-containing diet but consumed the control diet in preference.

Aggression↗
PubMed1975

Effects of cannabis resin on social behaviour in the laboratory mouse.

The effects of several dose levels of cannabis have been studied by an ethiological analysis of encounters between male mice injected with the drug and partners injected with Tween-saline. The duration of immobility was directly related to the logarithm of the dose of cannabis given. The ratio of flight in the treated animals to aggression in those mice with which they were paired showed a progressive increase as the dose of cannabis was raised from 4-50 mg/kg. A straight line relationship was found to exist between the logarithm of flight/aggression and logarithm of the dose of cannabis administered. Non-social activity and social investigation were decreased by the administration of cannabis; this was due to the time spent by the treated animals in immobility. Aggression was not significantly altered. Many mice given doses of 25-100 mg cannabis/kg showed an ataxic gait and the Straub tail reaction.

Aggression↗