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  • CHEMISTRY; METALLURGY
    • BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
      • MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
      • MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF (biocides, pest repellants or attractants, or plant growth regulators, containing micro-organisms, viruses, microbial fungi, enzymes, fermentates or substances produced by or extracted from micro-organisms or animal material A01N63/00; food compositions A21, A23; medicinal preparations A61K; chemical aspects of, or use of materials for, bandages, dressings, absorbent pads or surgical articles A61L; fertilisers C05); PROPAGATING, PRESERVING OR MAINTAINING MICRO-ORGANISMS (preservation of living parts of humans or animals A01N1/02); MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (micro-biological testing media C12Q)
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Bibliografia
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Keywords
MYOGLOBIN, MUSCLE, HYPOXIA, NITRIC OXIDE, FREE RADICALS, EFFICIENCY OF LOCOMOTION, MAGNETIC RESONANCE SPECTROSCOPY, XENON BINDING, PEROXIDASE ACTIVITY

The role of myoglobin in human muscle: relationship between molecular structure and function of the different isoforms

Università degli Studi di Milano
Abstract
As is well known, myoglobin (Mb) is a cytosolic metalloprotein found in relatively high concentration in the myocardium and in the skeletal muscle of man and other mammals. Its functional structure is that of a globin and a porphyrin ring containing an atom of iron (Fe), i.e. the heme. Fe is the linkage site of Mb with O 2 and other ligands of functional interest such as NO and its role, besides O 2 storage is that of a molecule facilitating O 2 diffusion from the periphery of the fibrocell to the site of O 2 utilization, i.e. the mitochondrion. Differently from other animals,in man Mb has five isoforms: Mb I (75-80% of the total: pI= 8.57); Mb II (15-20% of the total; pI= 7.29); Mb III, Mb IV e Mb V (in all together 5%; pI= 6.83) whose specific function is unknown. A privileged Mb ligand is nitric oxide (NO), which is known to inhibit reversibly cytochrome c oxidase (COX), the main regulator of cell respiration. The latter finding led Brunori (2001) to put forward the hypothesis that Mb at the level of the myocardium and of the skeletal muscle plays, besides the above indicated functions, that of a scavenger of intracellular NO thus preventing and /or reducing the NO-dependent COX inhibition thereby protecting the efficiency of the oxidative machinery.
The observation that Andean populations (Reynafarje, 1962) and high altitude Tibetan natives (Gelfi et al., 2004) are characterized by a greater concentration of muscle >>>

Principal Investigator
Paolo Cerretelli Università degli Studi di MILANO
Research Objectives
The aim of the present project is to verify the hypothesis put forward earlier by some of the applicants, that increased muscle myoglobin concentration, a possible scavenger of nitric oxide (NO) and O 2 free radicals, may improve the efficiency of oxidative phosphorylation in man thereby improving the economy of muscle exercise. The study shall be carried out at two different levels, a molecular one, assessing the affinity of Mb for ligands such as O 2, NO and H 2O 2 and an integrative one. This requires a close interaction among physicists, chemists, molecular biologist and physiologists thereby involving the experties different laboratories from four university departments and other scientific institutions.
As is well known, myoglobin (Mb) is a hemoprotein contained by human muscles at high concentration (0.5 mmol/kg of tissue) and, based on the large muscle mass of the body, in relatively large absolute quantity (10 mmol, i.e. about 200 grams). Mb is a small and densely packed protein consisting of a single polypeptide chain of 153 amino acid residues. It contains an iron-porphyrin heme group identical to that of hemoglobin and, like the latter, capable of reversible oxygenation and deoxygenation. There are five isoforms of human myoglobin : Mb I (75-80% of the total with isoelectric point = 8.57); Mb II (15-20% of the total pI= 7.29); Mb III, IV and V (together 5%, pI = 6.83). Besides the traditional function of >>>

Timescale
24 months
National and international background
As indicated before, Fe is the binding site of O 2 and of the other physiological ligands of myoglobin (Mb), such as CO, NO and reactive O 2 species. Five cavities surround the heme : the heme cavity and four cavities denoted by Xe1 to Xe 4 (Tilton et al. 1984). An important role in stabilizing the Fe-O 2 complex is played by distal histidine 64. Moreover, the presence of a number of aminoacidic residues that appear to be unchanged throughout the evolution, such as Leu 29, Val 168 and Ile 107, due to their hydrophobic effect in the cavities around the heme, are known to modulate the interaction of the Fe of the heme with the different ligands, according to various theories (“histidine gate”, Perutz and Metthews, 1966) and, more recently, a multipath process (Elber et al. 1990) as well as the ligand exit from the proximal site (Scott et al. 2001).

Here is a summary of the Mb functions:

1) Oxygen store and facilitated O 2 diffusion.
Mb plays an important physiological role in the delivery of O 2 in the cell (Wittemberg and Wittember, 1989) and, as an O 2 storage, stabilzes the O 2 flow within the muscle fiber throughout repeated contractions. Indeed, in the latter condition, blood flow is intermittent and O 2 flow to the mitochondrion (the “power plant”) is stabilized by the O 2 released from Mb. In order to carry out its >>>