Dr. Brajagopal Mujumder

Erwin Schrodinger first suggested that protein might vibrate. Scholars of the day are toiling best to establish his idea and hence to give a ringing endorsement. Their attempt is to link quantum mechanics with living organism. But it is hard to conduct experiment in vivo stage of mammalian body. Lot of attempts were undertaken to prove the hypothesis of protein vibration experimentally even in vitro stage if not in vivo stage of living organism. Since long, scientists held the view "Protein Vibrates like the string of a violin or the pipes of an organ" .  Moreover, they were of the opinion that these vibrations are incredibly high.

In spite of all these hurdles, scientists at different Universities have been trying to measure the vibration characteristics of protein. Recently, a team of scientists comprising of K.A. Niessen and others was able to conduct experiment successfully on protein vibration in vitro stage. They were able to measure the vibration spectrum of enzyme lysozyme - a protein that fights against bacteria by using Orientation Sensitive Terahertz near - field Microscopy system and observed that protein vibrates (rings) like a bell with a frequency in the range of a few terahertz or million - million terahertz and also held the view that vibration occurs in entire protein which ultimately lead to the phenomenon of transfer of energy across the protein 

This ultra - high - pitched vibration allows one protein long enough to interact with other proteins. This, as the scholars held, is due to the fact that biochemical reactions take place on a picoseconds time scale and since the vibration (ringing motion) lasts for only a few second, the scholars hypothesize that evolution has optimized enzymes to ring for the right amount of time". 

Practically speaking, vibrations in protein are occurred either due to the presence of electrostatic potential (force) in the cell or due to external stimuli. The magnitude of this electrostatic force lies in the range of (0.8 to 30) mv.  with respect to variable PH.  

In view of the above, it is relevant to raise the question - what roles do the protein vibration play with respect to living organism. These may be briefly stated as

(i) Based on molecular dynamic studies Iakhaiev and Iakhaiev  observed a molecular system in protein molecules which can co-ordinate and integrate structural changes in protein molecule. 

(ii)  Due to these tiny motions, proteins as held earlier are capable of binding with other molecules. This binding process is necessary for performing essential biological functions of living beings. Biological function of protein depends on its structural flexibility and changes thereof. Structural changes are critical with respect to cellular communication through membrane in channels to oxygen uptake along with delivery of hemoglobin. It is suggested by A.K. Niessen and others  that the vibrations which extend through protein play vital role in controlling these structured changes.

(iii) Recently, the author examined the role of vibration frequencies and vibration energies of protein in learning and memory formation with the application of EEG rhythms  (not included in the text). The author based on protein vibration approach also attempts to explain the in-depth principle of disaggregation of aggregated proteins due to resonance generated by native (resonator) and foreign protein vibration. For example, the author considered the role of disaggregation of aggregated proteins like alpha synuclein protein and tau protein by Heat Shock Protein -70 induced due to administration of the extract of Bacopa Monnieri (B.M) and, Ashwagandha etc of Indian Medicinal Herbs to the patients of Parkinson and Alzheimer . The author also analysed the modulating effects of Heat Shock Protein towards immune responses of cancerous Proteins  in the context of Protein Vibration. 

Based on these findings, it is held that conducting researches and studies on Protein vibration may open a new channel in therapeutic application of certain old age diseases.   

Parkinson's, Alzheimer's, Huntington's and other multiple neurodegenerative diseases are associated commonly with the process of memory loss and decline of cognition. These as the scholars held, are particularly due to accretion of aggregated protein and selective death of particular neural cells . 

No permanent cure for P.D. is still available. So far the drugs used are basically dopamine agonists and monoamines oxidize - B (MAO - B) inhibitors. However, these provide only symptomatic relief.

Considering the challenges that several natural products are being used for treating various ailments, Bacopa Monnieri (BM) commonly known as Brahmi - an Ayurvedic medicinal herb is, as the scholars held, found to act as anti-oxidant, anti-depressant, anti-inflammatory and anti microbial. It is now also regarded as the most neurotonic and memory boaster 
In India brahmi is used as a revitalizing herb by the Ayurvedic medical practitioners for almost 3000 years. It is small creeping herb. It belongs to scrophulariaceae family.  It has got number of branches, small oblong leaves and light purple. It grows in damp and marshy lands or sandy areas near streams in tropical regions. Apart from India, it grows in Nepal, Sri Lanka, China, Taiwan and Vietnam, it is also found in Florida and other Southern parts of USA. 

A team led by Aamir Zakir) conducted an experiment with caenorhabditis Elegans to examine the effect of B.M on parkinson's disease with the assumption that C. elegans many help in establishing an insights into therapeutic aspects of P.D. since their (60-80)% genes are homologous to human and it is also orthologs of P.D associated genes. The reason lies in the fact that dopaminergic neurodegeneration is induced by neurotoxins like 6-hydroxydopamine (6-OHDA) which in turn provides a pharmacological model for P.D. Basing on these facts, the team members conducted study on C. Elegans for examining the anti-parkinson's effect of neuron protective botanical species like B.M. However, other groups including the author also conducted studies on anti-parkinson's effect of B.M.

  It is an established fact that alpha synuclein protein in aggregated form that gets stiffened with reduction in energy is responsible for parkinson's disease. The molecular weight of this protein is 144.76 KD. This may be increased due to aggregation. 

The aggregation may follow self proteolytic activity of the protein within its full length of amino acids like.

(i) Protein of molecular weight 12.16KD with (14-133) amino acids.
(ii) Protein of molecular weight 10.44 KD with (40-140) amino acids formed through C-and N-terminal truncation.
(iii) Protein of molecular weights 7.27 KD with (72-140) amino acids formed through C-terminal.

It was observed by the scholars that 7.27 KD fragment gets aggregated even faster than a full length alpha-synuclein protein. Practically speaking, autoproteolytic products play a role as intermediates or co-factors in aggregation of alpha synuclein protein, This kind of aggregation of alpha synuclein protein is basically responsible for parkinson's disease.

Cure or symptomatic relief of the patient depends on the disaggregation of this alpha synuclein protein. It is expected that HSP-70 available (over expressed in presence of B.M) is able to undertake disaggregation of alpha synuclein protein. Now it is expected that fragment wise disaggregation in alpha synuclein protein may follow the process of self proteolytic activity as discussed. This means HSP-70 may disaggregate the aggregated alpha synuclein protein segment wise as in the case of aggregation. 

Under these conditions, an attempt has been taken here to evaluate frequency of vibrations and vibration energy thereof. These vibrations are expected to be generated due to electrostatic potential. An accurate amount of electrostatic potential at the surface of protein may be measured by considering the average potential over the surface of a protein which follows the contour of the dielectric discontinuity between protein interior and the solvent.
The magnitudes of this type of electrostatic potential is <30mV with respect to pH value. However, for the interest of the paper, the magnitude of electrostatic potential has been considered in between (0.8 to 1) mv with respect to variable pH value.
Vibration frequencies and energies of HSP 70, and alpha synuclein proteins have been calculated by considering electrostatic potential in two phases like 0.8 and 1 mv. based on normal equations of vibration frequency  & vibration .
It reveals from our calculations that both the vibration frequencies and vibration energy of Alpha synuclein protein are getting decreased with increase in molecular weight by the process of aggregation of different segments. But the vibration frequency and vibration energy of HSP is higher than those of alpha synuclein protein and also its different aggregated segments. Since HSP-70 is able to disaggregate the aggregated segments, alpha synuclein protein will gradually come to its original state. As a result, an amount of 29.87 KD will be reduced from the aggregated (175KD) mass of alpha synuclein protein in each cycle.

Thus our analytical approach is found to be in parity with the experimental findings where significant reduction in fluorescent intensities of aggregated alpha synuclein protein when worms (NL5901) were treated with BM as compared to untreated worms. So it is found that in both the cases, reduction in alpha synuclein protein constitutes the basic mechanism of treating PD.

From the above analysis, it reveals that HSP-70 by its characteristics functions as intra-cellular chaperones for aggregated alpha synuclein protein responsible for parkinson's disease. It is shown that HSP-70 helps in disaggregating the segment wise aggregation of alpha synuclein protein and thus helps to stabilize the protein conformation.

The present analysis supports the findings of Aamir Nazir and others that mother tincture (raw extract) of Bacopa monnieri (B.M) may decrease the aggregation of alpha synuclein protein and thereby may reduce its toxic out comes in the cells of C. elegans. Hence, identification and Isolation of proteins like HSP in Indian medicinal plants may take the lead in therapeutic treatment of a number of neurodegenerative diseases. Thus the studies conducted by the scholars with respect to anti-parkinsonian effect of B.M along with our analytic approach may open a new dimension regarding the utility of Indian system of Ayurvedic medicine.


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