Laser photolysis of liganded haem protein chains.

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The reactivities of the liganded states of the embryonic haemoglobins of the mouse with both O2 and CO were measured and compared with the reactivities of the adult protein.

Laser-photolysis experiments on the recombination of O2 with the partially oxygenated proteins indicates chain heterogeneity in the adult and embryonic EII and EIII species.

Gray RD. Quaternary structure of partially liganded intermediates of sheep carbon monoxide hemoglobin at alkaline pH.

J Biol Chem. Jan 25; (2)– Sawicki CA, Gibson QH. Quaternary conformational changes in human hemoglobin studied by laser photolysis of carboxyhemoglobin.

J Biol Chem. Mar 25; (6)–Cited by:   LASER PHOTOLYSIS STUDY OF CONFORMATIONAL CHANGE RATES FOR HEMOGLOBIN IN VISCOUS SOLUTIONS CHARLES A. SAWICKI AND MOHAMMAD A. KHALEQUE PhysicsDepartment,NorthDakotaStateUniversity,Fargo,NorthDakota ABSTRACT Rates for the R - T conformational change of deoxyhemoglobin formed by laser photolysis of carboxyhemoglobin Cited by:   The role of protein relaxation in Mb has been extensively investigated by laser photolysis because the photosensitivity of the complex of ferrous Mb with CO, O 2, and NO makes it possible to populate intermediate states and to follow the structural dynamics involved in the relaxation of the protein and the migration of the ligand through the Cited by:   The spectra do not support the earlier interpretation (Alpert, B., Banerjee, R.

and Lindqvist, L. () Proc.

Description Laser photolysis of liganded haem protein chains. FB2

Natl. Acad. Sci. U.S. 71, ) that this was due to a tertiary structural change of the protein. Introduction The photolability of liganded haem proteins has been utilised in many kinetic studies using flash by: Catherine Tetreau's 45 research works with citations and 2, reads, including: CO migration pathways in cytochrome P(cam) studied by molecular dynamics simulations.

Similarities in global properties of homopolymers and unfolded proteins provide approaches to mechanistic description of protein folding.

Here, hydrodynamic properties and relaxation rates of the unfolded state of carbonmonoxide-liganded cytochrome c (cyt-CO) have been measured using nuclear magnetic resonance and laser photolysis methods.

Hydrodynamic radius of the unfolded chain. The ligand-binding reaction in myoglobin (Mb) has been studied by a variety of techniques 1,2,3,4,5,6,7,8,9,10,11,12,13,Time-resolved and kinetic studies over wide ranges in time and.

Confirmation of Tertiary r in Liganded Quaternary T. As a control and to confirm our interpretation of r tertiary in T quaternary in the pulsed laser photolysis experiments of Viappiani et al.

(Fig. 2), we first carried out cw photolysis experiments on the T quaternary structure in the absence of allosteric effectors (T −) (Fig. 4).In these experiments, the sample is exposed to the cw.

The photolysis was produced by a laser flash at nm of total duration 50 nsec and the product was observed during a further nsec. Measurements were performed on carboxy and oxy derivatives of human hemoglobin, those of the valency hybrids (4), isolated chains (~ and ~), on free carboxyheme and also on free hemoglobin and methemoglobin.

Fig. TR 3 spectra of ferrous Mb in interaction with NO and kinetics of the ν Fe-His mode. (A) Ground-state Raman spectra of deoxy Mb and of six-coordinate NO-liganded Mb (-5 ps), and difference TR 3 spectra after NO photodissociation at indicated time delays after photolysis.(B) Evolution of normalized area of the ν Fe-His mode contour (– cm-1, between the vertical dotted lines in.

The oxygen reaction of the fully reduced respiratory chain in membranes from oxygen-limited Escherichia coli was studied at sub-zero temperatures usin. On the millisecond time scale following photolysis, CO diffuses out of the haem pocket and into the solvent. Partial photolysis time courses were collected at decreasing levels of breakdown.

Laser power was reduced until ⩽10% breakdown of the fully liganded Hb was obtained. A nanosecond laser flash-photolysis technique was used to study bimolecular and geminate molecular oxygen (O2) rebinding to tetrameric human hemoglobin and its isolated α and β chains in buffer.

CONCLUSIONS Laser photolysis of methylisocyanide with the R and T states of carp hemoglobin reveals small, but definite, differences in geminate recombination yields and kinetics. The geminate process is primarily controlled by the tertiary structure of the ligand binding site, which apparently does not differ much in the two liganded.

Simplified kinetic scheme depicting events following photolysis of liganded Mb (A). The photolysed ligand (O 2, CO, NO, and others) populates the so‐called distal pocket (primary docking site, B), and thereafter may: (i) diffuse out into the solvent (S) via the His‐gate, 26 the most probable path according to Scott et al.

27; (ii) rebind to the metal by an intramolecular geminate process B. The rapid changes in absorbance observed at the isosbestic points of unliganded and liganded hemoglobin following laser photolysis provided a value of. The utility of this method is exemplified on a dataset of protein chains from the globin family and a dataset of tetrameric hemoglobins.

long enough after laser photolysis to enhance the. Proc. Nat. Acad.

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Sci. USA Vol. 71, No. 2, pp.February TheKinetics ofConformational Changesin Hemoglobin, Studied by Laser Photolysis (myoglobin/cooperative interactions/ligand binding) B. ALPERT*, R. BANERJEE*, AND L.

LINDQVISTt * Institut deBiologie Physico-chimique, 13, rue Pierre et Marie Curie, Paris, France; and t Laboratoire de Photophysique. The photolysis of HbO2 and HbCO has been investigated with picosecond laser techniques.

Transient absorption spectra were measured in the Soret and visible regions after excitation with or nm pulses. The photoproducts appeared within 8 psec and exhibited considerably broadened deoxyhemoglobin-like spectra, which persisted to psec. Although commendable progress has been made in the understanding of the physics of protein folding, a key unresolved issue is whether Kramers’ diffusion model of chemical reactions is generally applicable to activated barrier crossing events during folding.

To examine the solvent viscosity effect on the folding transition of native-like trapped intermediates, laser flash photolysis has been. Rate constants for the bimolecular recombination of CO were determined by partial laser flash photolysis by reducing laser power.

Geminate rebinding from within the haem pocket is complete by t > ns, after which partial photolysis produces a fraction of unliganded haems that composes 10–20% of the total haems, thus making the most.

A chemically modified form of cytochrome c (cyt. c), termed carboxymethyl cytochrome c (cm cyt. c), possesses a vacant sixth coordination site to the haem iron that is available to bind external ligands.

We present data on the rapid flash photolysis of CO from the ferrous haem iron of cm cyt. c and describe the kinetics and spectral transitions that accompany the recombination. This was. Both time-resolved laser flash photolysis studies and ESR spectroscopic investigations clearly demonstrate the formation of C+) using PET conditions.

Upon addition of H-donors such as methanol the signal of C+) is quenched as monitored both by laser flash photolysis. Beta globin (also referred to as HBB, β-globin, haemoglobin beta, hemoglobin beta, or preferably haemoglobin subunit beta) is a globin protein, which along with alpha globin (), makes up the most common form of haemoglobin in adult humans, the HbA.

It is amino acids long and has a molecular weight of 15, adult human HbA is a heterotetramer consisting of two alpha chains and.

The urea, guanidine hydrochloride, salt, and temperature dependence of the rate of dissociation of CO from a nonequilibrium state of CO-bound native ferrocytochrome c has been studied at pH 7.

The heme iron of ferrocytochrome c in the presence of denaturing concentrations of guanidine hydrochloride (GdnHCl) and urea prepared in M phosphate, pH 7, binds CO. When the unfolded protein.

The urea, guanidine hydrochloride, salt, and temperature dependence of the rate of dissociation of CO from a nonequilibrium state of CO-bound native ferrocytochrome c has been studied at pH 7.

Details Laser photolysis of liganded haem protein chains. EPUB

The heme iron of ferrocytochrome c in the presence of denaturing concentrations of guanidine hydrochloride (GdnHCl) and urea prepared in M phosphate, pH 7, binds CO.

Before binding to haem a 3, CO and O 2 bind to CuB intermediately 6,7; the reverse reaction, involving ligand transfer from haem a 3 out of the protein via CuB, can be studied using flash photolysis. Myoglobin, a small globular haem protein that binds gaseous ligands such as O2, CO and NO reversibly at the haem iron, serves as a model for studying structural and dynamic aspects of protein.

Time courses for full photolysis (panel C) and partial photolysis (11% photolysis for both; panel D) of O 2 from HbA and α-cpβ. Experiments were conducted in air-equilibrated M potassium phosphate and 1 mM EDTA, pH at 20 °C.

The concentrations of heme in the HbA and α-cpβ samples were 73 and 56 μM, respectively. plexes of cytochrome d [ and other haem pro- teins [14] has been reported. However, photolysis at this temperature did give rise to a spectrally distinct form of the oxidase in the presence of O2 [9,11], sug- gesting photolysis of the CO compound and ligand exchange.Fragment rotational population distributions for OH and OD were observed by laser-excitation fluorescence following the nm photolysis in a free jet molecular beam of HOCl and DOCl, respectively.

The photodissociation dynamics show a strong preference for the Π(A′) OH or OD lambda doublet component in both spin–.

Dissociation of the CO from the iron generates a characteristic five-coordinate deoxyhaem absorbance spectrum, which is then replaced by a six-coordinate spectrum as the protein folds and the native methionine ligand reattaches to the haem.

We used laser photolysis to trigger the folding, and we monitored its progress through transient.