Project proposal
In vitro analysis of fifteen antidiabetic plants for
-glucosidase inhibition
Submitted to
RESEARCH SOCIETY FOR THE STUDY OF DIABETES IN INDIA
Principal Investigator
Dr. Kavitha Thirumurugan
School of Bio Sciences and Technology
VIT University, Vellore - 632 014
Tamil Nadu, India.
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Title: In vitro analysis of fifteen antidiabetic plants for -glucosidase inhibition
Name and address of the Principal Investigator:
Dr. Kavitha Thirumurugan
207, Structural Biology Lab
Centre for Biomedical Research (CBMR)
School of Bio-Sciences & Technology (SBST)
VIT University, Vellore-632 014
Email: kavithiru@hotmail.com
Introduction
Type 2 Diabetes Mellitus (Type 2 DM) is an endocrine characterized by
hyperglycemia due to insulin resistance and
insulin deficiency resulting from β-cell
dysfunction. DM is associated with a variety of symptoms like polyphagia, polyurea and
polydipsia. Major chronic complications include accelerated macro-vascular diseases like
retinopathy, renal disease and neuropathy. Hyperglycemia leads to increased aldolase
reductase activity and accumulation of sorbitol, accelerated non- enzymatic glycosylation of
proteins and increased diacyl glycerol levels.
There is a steady rise in the rate of incidence of Type 2 DM in the world. According
to WHO (2010), 33 million cases of diabetes were reported in the US in the year 2000 and
the number is estimated to rise to 66.8 million by 2030. In Europe, 33.3 million cases were
reported in the year 2000 and it is estimated to rise to about 48 million by 2030. In India, 19.3
million cases were reported in the year 1995 and it is estimated to rise to about 57.2 by 2025.
In India, it is becoming a killer disease next to coronary heart disease. The causes could be
attributed to sedentary lifestyle, lack of physical exercise and obesity.
Various systems of medicines like Ayurveda, Unani and Siddha are practised all over
the world. In India, Ayurveda is an ancient practise that evolved over 5,000 years ago.
According to World Health Organisation, plant derived drugs constitute the mainstay of
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nearly 80 % of the population for their primary health care. In this study, we have screened
fifteen antidiabetic plants for their potential to inhibit α-glucosidase. These plants are:
Aconitum heterophyllum, Acorus calamus, Clerodendron serratum, Cyperus rotundus,
Marsdenia tenacissima, Messua ferrea, Nigella sativa, Picrorhiza kurroa, Piper
retrofractum, Plumbago zeylanica, Rubia cordifolia, Saussurea lappa, Symplocos racemosa,
Terminalia arjuna, Zingiber officinale.
α-glucosidase is a membrane bound enzyme which is located on the brush border of
the small intestine and it is required for the break down of complex carbohydrates into
monosaccharides that can be absorbed. The α-glucosidase inhibitors (AGI) inhibit the
catalytic activity of the enzyme and thereby delay the absorption of ingested carbohydrates
but do not prevent it. Only monosaccharides such as glucose and fructose can be transported
out of the intestinal lumen into the blood stream, in the presence of AGI. This results in the
reduction of postprandial glucose and insulin peaks. Flavonoids, polyphenols and glycosides
are found to be effective AGI’s. Few commercially available AGI’s : acarbose and voglibose
are competitive inhibitors of the enzyme. However, these oral hypoglycaemic agents have
prominent side effects and failed to alter the course of diabetic complications. Therefore, we
have to look for alternative herbal medicines that have fewer side effects for the treatment of
diabetes.
In this study we have to find the potential plants with maximum inhibition for alpha
glucosidase enzyme. These plants will be included for kinetic analysis to know the mode of
inhibition. Following this, the best performing plant will be evaluated for its postprandial,
anti-hyperglycemic activity using animal models. Next, we will perform bioactivity guided
fractionation to obtain pure, active fraction and the identity of the compound will be known
from the mass spectrometry. After this, structural and functional characterization of the
compound will be performed. We hope the active antidiabetic compound derived from plant
source can be given to human volunteers for future epidemiological study.
Review of Literature
Aconitum heterophyllum (AH), Acorus calamus (AC), Clerodendron serratum (CS), Cyperus
rotundus (CR), Marsdenia tenacissima (MT), Mesua ferrea (MF), Nigella sativa (NS),
Picrorhiza kurroa (PK), Piper retrofractum (PR), Plumbago zeylanica (PZ), Rubia cordifolia
(RC), Saussurea lappa (SL), Symplocos racemosa (SR), Terminalia arjuna (TA), Zingiber
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officinale (ZO) are included in the current study. Plant parts are purchased from an Ayurvedic
shop, Vellore and ground to yield a powdered form to be used for the solvent extraction.
Aconitum heterophyllum belongs to the family Ranunculaceae. The plant is commonly found
in alpine and sub- alpine region of the Himalayas at altitudes between 1,800-4,500 m. The
roots of the plant have medicinal properties. Nisar et al. [1] extracted various
butyrylcholinestrase and acetylcholinesterase inhibitors from the roots of the plant. Ahmed et
al. [2] extracted norterpenoid alkaloids from the roots of these plants having antibacterial
activity. A study conducted by Atal et al. [3] showed that the ethanolic extracts of this plant
stimulates phagocytic function while inhibiting the humoral component of the body’s
immune system, thus acting as an immunomodulator.
Acorus calamus belongs to the family Araceae. Belska et al. [4] showed that a pectic
polysaccharide obtained from this plant activates macrophages and stimulate Th1 response.
Jain et al. [5] reported that the ethanolic extract of the leaves promoted wound- healing
activity in rats. Hu et al. [6] indicated inhibitory effects of the aqueous extract on waterbloom forming species of algae. A study conducted by Si et al. [7] showed that ethyl acetate
extract had insulin releasing and α- glucosidase inhibitory activity. The IC
50
value reported
by the authors was 0.41 μg/ml.
Clerodendron serratum belongs to the family Verbenaceae. Vidya et al. [8] showed that the
ethanolic extract of the roots of Clerodendron has hepatoprotective activity against carbon
tetrachloride induced toxicity in rats. Narayanan et al. [9] showed that alcoholic extract of the
roots of this plant has anti-inflammatory activity. It was also shown to have antinociceptive
and antipyretic activity in animal model.
Cyperus rotundus belongs to the family Cyperaceae. The rhizome of this plant has been
reported by Lee et al. [10] to play a major role in the protection of neurodegenerative
disorders due to its antioxidant and free radical scavenging activity. Pal et al. [11] showed
the ethanolic extract of this plant having analgesic properties. Soltan et al. [12] reported
hydro-alcoholic extract of this plant displaying antiviral effect against Herpes Simplex.
Kilani- Jaziri et al. [13] showed ethyl acetate extracts of Cyperus to inhibit xanthine oxidase
in XO induced human chorionic myelogenous leukemia cells by H2O2. Kilani et al. [14]
mentioned significant antibacterial effect of Cyperus. Raut et al. [15] reported hydro-
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ethanolic extract of Cyperus to reduce blood glucose level significantly in alloxan induced
diabetic rats.
Marsdenia tenacissima belongs to the family Asclepiadaceae. Qian et al. [16] showed the
significant antitumor effects of this herb in experimental and clinical applications. Xia et al.
[17] isolated novel glycosides from the root of this plant. Hu et al. [18] showed the efficiency
of tenacigenin derived from this plant to reverse multidrug resistance in cancerous cells.
Mesua ferrea or Nagakesara is a medium- sized to large evergreen tree from the family of
Clusiaceae. Mazumder et al. [19] studied the antibacterial properties of the flowers of this
plant. Meherji et al [20] reported estrogenic and progestational activity of this plant on mice
and humans. Recently it has been shown that calophyllolide isolated from Mesua is effective
in reducing the increased capillary permeability (induced in mice by iiistamine, 5-HT and
bradykinin). Main use of stamen has been described to control bleeding in menorrhagia and
piles. Xanthones, a number of 4-phenylcoumarin derivatives, friedelin and triterpenes have
been isolated from the plant. Xanthones are isolated from the heartwood; coumarin
derivatives from the seeds; canophylial, canophyliol and canophyllic acid from the leaves.
Recently, a tetraoxygenated xanthone was isolated from the heartwood and bark of the plant.
Fatty acid composition of the seed oil has also been studied using several methods.
Nigella sativa belongs to the family Ranunculaceae. It is an annual herb which has numerous
medicinal properties and thus popularly used in folk medicine. According to Geng et al. [21],
the volatile oil extracted by hydro-distillation contains thymoquinone (3.8 %) which is
involved in anti-inflammatory activities in vivo and in vitro. Banerjee et al. [22] proved the
therapeutic potential of thymoquinone in pancreatic cancer. Isik et al. [23] showed its
potential adjuvant effects to improve immunotherapy in the treatment of allegic Rhinitis.
Fixed oil and water extract of this plant (0.1% v/v) has shown to considerably reduce
formation of sickle cells due to its calcium antagonistic and antioxidant activities [24]. Oral
administration of the ethanol extract of N. sativa seeds to diabetic rats administered with
streptozotocin reduced hyperglycemia [25]. According to Arayne et al. [26], methanolic
extract of Nigella significantly inhibits glucose utilisation in the intestine of rats. Petroleum
ether extract of the seeds of this plant has shown to exert insulin sensitising action. Kanter et
al. [27] showed that N. sativa treatment in streptozotocin-induced diabetes in rats,
significantly reduced lipid peroxidation and serum nitric oxide levels by increasing
antioxidant enzyme activity. Abdel zaher et al. [28] showed that the oil extracted from the
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plant blocks nitric oxide overproduction and ceases morphine-induced tolerance and
dependence in mice. Al-nagger et al. [29] showed neuropharmacological activity of the
methanolic extracts of this plant. It was showed that it possess potent CNS depressant and
analgesic activity.
Picrorhiza kurroa belongs to the family Scrofulariaceae. Verma et al. [30] found that a
glycoside namely picroliv is a hepatoprotective compound. The compound is also shown to
have cholerectic and anti-cholestatic effects in rats and guinea pigs. It also has anti-viral and
immuno-modulatory compound. Yadav et al. [31] showed that picroliv restored cadmium
induced abnormalities in the liver of male rats. Anand et al. [32] showed that picroliv has
anti- inflammatory and anti-carcinogenic properties. Dhuley et al. [33] showed that picroliv
has antioxidant effects. Banerjee et al. [34] explored the healing potential of the methanolic
extract of the plant in indomethacin induced stomach ulcers in mice. Khajuria et al. [35]
explored the potential of the compounds derived from this plant as alternative adjuvant.
Zhang et al. [36] derived tannins from the plant that could inhibit cyclooxygenase and lipid
peroxidation.
Piper retrofractum belongs to the family Piperaceae. Hardik et al. [37] showed hexane and
methanol extracts of the plant having potent antileishmanial activity. Komalamisra et al. [38]
reported the ethanolic extract of the plant having larvicidal and insecticidal properties.
Limyati et al. [39] suggested that the fruits of this plant has potent anti- bacterial and antifungal properties. Nakatani et al. [40] found several phenolic amides from the plant which
might have potent antioxidant properties.
Plumbago zeylanica belongs to the family Plumbaginaceae. Chen et al. [41] showed
plumbagin isolated from this plant possessing significant anticancer activity. Maniafu et al.
[42] showed that hexane and chloroform extract of this plant had significant larvicidal
activity. Edwin et al. [43] showed that the acetone and ethanolic extracts of the leaves of this
plant had reversible concentration dependent oestrogenic and anti-oestrogenic activity.
Checker et al. [44] showed that plumbagin isolated from this plant has significant antiinflammatory effects.
Rubia cordifolia belongs to the family Rubiaceae. Lu et al. [45] successfully isolated antioxidative constituents from ethyl acetate extract of this plant. Patil et al. [46] showed that
alcoholic extract of this plant increased brain gamma-amino-n–butyric acid levels and
decreased brain dopamine and plasma corticosterone levels. They also showed that the
6
extract inhibited acidity and ulcer formation. They also showed that the extract decreased
blood sugar level that was increased by alloxan treated animals.
Saussurea lappa belongs to the family Asteraceae. Yaeesh et al. [47] proved antihepatotoxic
activity of the aqueous- methanol root extract of this plant in mice. Yu et al. [48] showed that
ethanol extract of this plant inhibited Streptococcus mutans in a dose dependent manner.
Gilani et al. [49] showed that the plant contains cholinergic and calcium antagonist
ingredients which is helpful for use in constipation and spasms. Sarwar et al. [50] studied the
effect of ethanolic extracts on lymphocyte proliferation. Rao et al. [51] isolated antifungal
constituents from the roots of the plant. Kim et al. [52] proved the anti-tumour properties of
the plant.
Symplocos racemosa belongs to the family Symplocaceae. Miszczak-Zaborska et al. [53]
reported glycosides isolated from this plant to inhibit thymidine phosphorylase whose
overexpression is linked to angiogenesis. Ahmad et al. [54] studied the kinetics of an
inhibitor of α-chymotrypsin. Lodhi et al. [55] studied the kinetics of triaconityl palmitate
which is a urease inhibitor. In a phytochemical investigation, Ahmad et al. [56] found new
phenolic glycosides of salirepin series in the n-butanol fraction of the bark of S. racemosa.
Abbasi et al. [57] found ethyl substituted glycoside that inhibits lipoxygenase. Choudhary et
al. [58] found phenolic glycosides that inhibited human nucleotide pyrophosphatase
phosphodiesterase.
Terminalia arjuna belongs to the family Combretaceae. It is common throughout India
especially in the sub-Himalayan tracts and Eastern India. This plant is widely known to prove
comprehensive relief to the people suffering from cardio-vascular diseases, especially
hyperlipidemia and ischemic heart disease. Some important findings related to the above
mentioned activity has been studied by Mahmood et al. [59]. Halder et al. [60] studied the
anti- inflammatory, immunomodulatory and antinociceptive activity of the bark in mice and
rats. Kumar et al. [61] showed that T. arjuna bark extract attenuated catecholamine- induced
myocardial fibrosis and oxidative stress. Khan et al. [62] showed its antimicrobial activity
against multi drug resiastant (MDR) strains of fungi and bacteria of clinical origin. Alam et
al. [63] isolated oleanane-type triterpene glycosides which suppresses the release of nitric
oxide and superoxide from macrophages and also inhibited aggregation of platelets. Reddy et
al. [64] studied the effect of T. arjuna extract on adriamycin–induced micronuclei formation
in cultured human peripheral blood lymphocytes. Dwivedi et al. [65] showed that Terminalia
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arjuna has been found to be useful in diabetes associated with ischemic heart disease. In our
current in vitro study, T. arjuna significantly inhibits α-glucosidase due to the presence of
glycoside.
Zingiber officinale belongs to the family Zingiberaceae. It is commonly known as ginger.
Wang et al. [66] showed anti-microbial and antioxidant effects of the fractions obtained from
the rhizome of this plant. Takahashi et al. [67] reported that a formulation of the plant was
effective in controlling alcohol hangover symptoms. Incharoen et al. [68] showed that dried
fermented ginger improved intestinal function. Khan et al. [69] showed that ethanolic extract
of this plant had remarkable inhibitory activity against multi-drug resistant bacterial and
fungal strains. Lin et al. [70] showed that the plant has larvicidal constituents. Wang et al.
[71] proved the anti-invasive property of the constituents of the plant in hepatocarcinoma
cells. A work carried out by Nojpha et al. [72] showed that ginger promoted glucose transport
in muscle cell line. Khushtar et al. [73] explored the protective action of ginger oil on gastric
ulcer induced by aspirin in rats. Shanmugam et al. [74] studied protective effect of ethanolic
extract of ginger on antioxidant enzymes in rats and its effectiveness against renal damage
induced by alcohol. Akhani et al. [75] showed that aqueous extract of the rhizome produced
significant increase in insulin levels and decrease in fasting glucose levels in diabetic rats.
Treatment with the extract also caused reduction in serum cholesterol levels, blood pressure
and serum triglycerides in diabetic rats.
Aims
1. To measure the -glucosidase inhibitory activity of the fifteen medicinal plants
2. To perform kinetic analysis on the potential plants
3. To isolate active compound from the best plant
4. To determine the structure of the compound.
5. To assess the level of safety, toxicity of the isolated compound.
Work plan (including detailed methodology and time schedule)
Plant extracts and standard
Methanolic extracts of the plant parts will be prepared using a soxhlet apparatus. The extracts
will then fed into a rotary evaporator to remove the solvent (methanol) and the dried extract
8
obtained will be stored at –5 ⁰C. The standard voglibose will be purchased and ground. It is
dissolved in distilled water and centrifuged at 6000 rpm. The supernatant is taken and
appropriately diluted.
Reagents, plastic ware and instrumentation
The reagents used for the enzyme assay will be purchased from SISCO research laboratories
Pvt. Ltd.– Mumbai. The glasswares including Soxhlet apparatus was purchased from Borosil
Glass works Ltd.- Mumbai. The plastic wares will be purchased from Tarsons products Pvt.
Ltd.- Kolkata. The 96-well plate reader has to be purchased from Bio Tek USA Inc.
In vitro analysis of -glucosidase inhibitory activity
Modified Pistia-Brueggeman's method is used for this spectrophotometric kinetic end-point
assay [76]. A solution of -glucosidase has been prepared at a concentration of 1 U/ml at pH
6.8 with 50 mM phosphate buffer. 1 mM p-nitrophenyl--glucopyranoside (PNPG) is
prepared at pH 6.8 using phosphate buffer. The plant extracts are dissolved in 50 mM
phosphate buffer at pH 6.8. Inhibitor concentration ranging from 0.1 μg/ml to 1 mg/ml has
been prepared. In a 96 well plate, 50 l of 50 mM phosphate buffer, 20 l of extract or
control (buffer or inhibitor control) and 10 l of enzyme are mixed and shaken on a plate
shaker. After incubation for 5 minutes at 37 °C, 20 l of substrate is added to appropriate
wells and shaken, in order to commence the reaction. The plate is incubated for 30 minutes at
37°C and 50 l of 0.1 M sodium carbonate will be added to terminate the reaction and ionize
the p-nitrophenol, if formed. The yellow colour produced will be quantitated by colorimetric
analysis, by reading the absorbance at 405 nm in a 96 well plate reader. Each sample will be
performed in triplicate, along with appropriate blanks. The % inhibition is obtained using the
formula:
% inhibiton = {Absorbance(control) –Absorbance(sample)}/ Absorbance(control)
IC50 value can be defined as the concentration of extract inhibiting 50% of -glucosidase
under the stated assay conditions. In case of significant inhibition, IC50 are determined by
nonlinear regression by fitting to a sigmoidal dose-response equation with variable slope. All
values are represented as Mean  Standard Deviation.
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Kinetics of -glucosidase inhibition by Methanolic plant extracts
Inhibition mode of the extracts against -glucosidase activity will be measured with
increasing concentrations of PNPG (0.125, 0.25, 0.5 and 1mM) as a substrate in the absence
or presence of the plant extracts at two different concentrations for each plant extract.
Optimal doses of the plant extracts are determined based on the results from inhibitory
activity assay as described earlier. Inhibition type for the plant extracts is determined by
Lineweaver–Burk plot analysis of the data, which were calculated from the results according
to Michaelis-Menten kinetics [77, 78]. Experimental inhibitor constant (Ki) values were
determined by Double reciprocal plots. The theoretical value of Ki is obtained using the
formula :
Ki = Vmax‫* ׳‬I/( Vm-Vmax‫)׳‬
Conclusion
Diabetes mellitus is a progressive metabolic disorder affecting majority of the population
across the world. There are various measures to manage and treat this killer disease. The
main effect of diabetes is increase in glycemic level. To reach normoglycemic level, along
with
insulin
other
oral
hypoglycemic
agents
like
sulfonylureas,
biguanides,
Thiazolidinediones (TZD), α-glucosidase inhibitors (AGI) and incretin mimetics (GLP-1,
GIP, DPP-4 inhibitors) are used. α-glucosidase inhibitors delay the action of α-glucosidases
to break complex carbohydrtaes in to simple sugars, thereby lowering the absorption of
glucose. These inhibitors play a vital role in reducing the postprandial hyperglycemia. As a
consequence of their pharmacological action, α-glucosidase inhibitors also cause a
concomitant decrease in postprandial plasma insulin and gastric inhibitory polypeptide and a
rise in late postprandial plasma glucagon-like peptide 1 levels. In individuals with normal or
impaired glucose tolerance with hyperinsulinemia, α-glucosidase inhibitors decrease
hyperinsulinemia and improve insulin sensitivity [80]. Postprandial hyperglycemia
contributes to raise in glycated hemoglobin (HbA1c) level, which as an indicator of total
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glycemic load, is tightly correlated with the incidence of micro- and macroangiopathy in
Type 2 diabetes. It can induce or deteriorate fasting hyperglycemia and be associated with
coagulation activation and/or lipid metabolism abnormalities [81]. It has been reported that αglucosidase inhibitors had a beneficial effect on glycated haemoglobin [82]. Epidemiological
data from the United Kingdom Prospective Diabetes Study (UKPDS) also showed that there
is a 14–16% decrease in macrovascular complications for every 1% absolute reduction in
glycated haemoglobin [83]. α-glucosidase inhibitors like acarbose, miglitol and voglibose are
used in conjunction with other anti-diabetic drugs. But these inhibitors have some side effects
like flatulence and diarrhea. This indicates that newer AGI’s with lesser side-effects needs to
be discovered. Therefore, we have to screen potential antidiabetic plants for α-glucosidase
inhibition.
Time Schedule of activity
Activity block
Time required
1. Crude extract: Inhibition assay for α-glucosidase activity and
3 months
kinetic analysis
2. Testing the effect of extract on animals
3 months
3. Fractionation of extract, active compound isolation
6 months
4. Identification and structure determination of the compound
3 months
Total = 15 months
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Budget
Details of financial requirements for three years (with justifications) and phasing for
each year:
S.No. Head
3 months
9 months
15 months
Total
1.
Consumables
50,000
25,000
25,000
1,00,000
2.
Travel (within
India)
-
-
10,000
10,000
Total =
1,10,000
Justification of the Cost
1. Consumables
Chemicals including biochemical and enzymatic test kits, reagents and glasswares
have to be purchased for the proposed study. High purity chemicals are required to get
reproducible results in enzyme assays. Quality kits are essential to get reliable results with
plasma enzyme assessments.
2. Travel
To determine the structure of the active compound, NMR studies have to be carried at
the Indian Institute of Science, Bangalore. Also, SAIF facility at IIT, Chennai will also be
required to use GC-MS. Project scholars have to visit these institutes to perform the research.
To communicate the results of our research, we need to present the data at
national/international conferences. Hence, the grant towards the travel purpose is justified.
Whether the project is being partly funded from any other source?
No
12
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Detailed Biodata
1. Name of the Applicant:
Dr. Kavitha Thirumurugan
2. Mailing Address (Indicate Telephone, Fax, Email, etc)
207, Structural Biology Lab
Centre for Biomedical Research
School of Bio-Sciences & Technology
VIT University, Vellore-632 014, INDIA
Email: kavithiru@hotmail.com
Fax: +91-416-2243092
Tel: +91-416-2202510
3. Date of Birth & Gender: June 2, 1971 Female
4. Educational Qualifications (Starting from Graduation onwards). Details of the
degree/diploma, university, year of passing, subject of specialization, class/grades
obtained etc.
Name of the
University
Degree
B.Sc
Year of
Field of Study
OGPA
1992
Agriculture
3.94/4.00
1995
Plant Breeding
9.40/10.00
passing
Tamil Nadu
Agricultural
University,
Coimbatore
M.Sc
Tamil Nadu
Agricultural
& Genetics
University,
Coimbatore
24
Ph.D
Tamil Nadu
1998
Agricultural
Plant Breeding
9.66/10.00
& Genetics
University,
Coimbatore
Advanced
Birkbeck College,
2006-2007
Techniques in
Certificate
University of
Structural
Course
London, UK
Molecular
Distinction
Biology
5. A. Details of professional training and research experience, specifying period.
Professional research experience as a Postdoctoral Researcher at the University of
Leeds will be a valuable tool to tap the research potential of my current research students. I
had investigated the interaction between actin and myosin using millisecond rapid mixing
kinetics apparatus to capture the transient conformational states of the complex. This will
enable me to guide my students on the kinetics of enzyme-substrate complex. I had
performed NADH-coupled ATPase activity, baculovirus expression, and protein purification
using various chromatography tools. To study the structure of myosin, I had intensively used
electron microscopy and image processing. This will assist me to observe the histological
sections of pancreatic tissues and appreciate its relevance in the context of current research on
diabetes. Professional teaching experience as an Associate Professor (Biochemistry) at VIT
University is tremendously helpful to translate theoretical knowledge into practical output.
B. Details of employment:
Sl No.
Institution
Position
From (Date)
To (date)
25
Place
1.
VIT University, Vellore
Associate Professor
21.10.08
Present
2.
University of Leeds, UK
Post-doctoral
06.06.01
20.10.08
1999
2000
Research Fellow
3.
Adhi Parasakthi Agricultural
Assistant Professor
College, Kalavai
C. List of significant publications during last five years (with details)
Publications (Numbers only): 24

Research Papers (peer-reviewed): 8

Conference papers (peer-reviewed): 8

Research papers (National): 8
Research Papers (Peer-reviewed) (available in Pubmed):
1.
Houmeida A., A. Baron, J. Keen, N. Khan, P. J. Knight, W. F. Stafford III, K.
Thirumurugan, B. Thompson, L. Tskhovrebova, J. Trinick (2008) Evidence for the
Oligomeric State of ‘Elastic’ Titin in Muscle Sarcomeres. J. Mol Biol 384(2): 299312
2.
Sellers, J. R., K. Thirumurugan, T. Sakamoto, J. A. Hammer III, P. J. Knight (2008)
Calcium and cargoes as regulators of myosin 5a activity. Biochem Biophys Res
Commun. 369(1): 176-81
3.
Kovacs, M., K. Thirumurugan, P. J. Knight, J. R. Sellers (2007) Load-dependent
mechanism of nonmuscle myosin 2. PNAS 104(24): 9994-9
26
4.
Thirumurugan, K., T. Sakamoto, John A Hammer III, J. R. Sellers, P. J. Knight
(2006) The cargo-binding domain regulates structure and activity of myosin 5. Nature
442 (7099): 212-215
5.
Knight, P. J., K. Thirumurugan, Y. Yu, F. Wang, A.P. Kalverda, W.F. Stafford, J.R.
Sellers, M. Peckham (2005) The predicted coiled-coil domain of myosin 10 forms a
novel elongated domain that lengthens the head. J. Biol. Chem. 280 (41): 34702-3470
6.
Wang*, F., K. Thirumurugan*, W.F. Stafford, J.A. Hammer, P.J. Knight, J.R.
Sellers (2004) Regulated conformation of Myosin V. J. Biol. Chem. 279, 2333-2336.
(*Equal contribution)
7.
Burgess, S. A., M.L. Walker, K. Thirumurugan, J. Trinick, P.J. Knight (2004) Use
of negative stain and single-particle image processing to explore dynamic properties
of flexible macromolecules. J. Struct. Biol. 147, 247-258
8.
White, H. D., K. Thirumurugan, M.L. Walker, J. Trinick (2003) A second
generation apparatus for time-resolved electron cryo-microscopy using stepper
motors and electrospray. J. Struct. Biol. 144, 246-252.
Conference Papers presented in National/International Conferences
1. Thirumurugan, K., S. A. Burgess, Fang Zhang, J. R. Sellers, P. J. Knight (2009)
Cryo-Electron Microscopy of Myosin 5 on actin. 2009 Biophysical Society Meeting
Abstracts. Biophysical Journal, Supplement, Abstract. Page
2. Thirumurugan, K., E. Forgacs, T. Sakamoto, H. D. White, P. J. Knight (2008)
Structural basis for gated release of ADP from Myosin 5 on actin. 2008 Biophysical
Society Meeting Abstracts. Biophysical Journal, Supplement, Abstract, 2255-Pos.
Page 453
27
3. Thirumurugan, K., P. J. Knight, S. A. Burgess, T. Sakamoto, J. A. Hammer, J. R.
Sellers, (2006) Folding and ATPase regulation of myosin 5. 2006 Biophysical Society
Meeting Abstracts. Biophysical Journal, Supplement, Abstract, 2083-Pos.
4. Thirumurugan, K., S. A. Burgess, J. Trinick, H. D. White, P. J. Knight, (2005) Twoheaded binding of muscle myosin 2 to F-actin in ATP. 2005 Biophysical Society
Meeting Abstracts. Biophysical Journal, Supplement, 18a, Abstract, 92-Plat.
5. Thirumurugan, K., P. J. Knight, J. A. Hammer, J. R. Sellers, (2005) The globular
tail domain regulates the enzymatic activity of myosin V. 2005 American Society for
Cell Biology Meeting Abstracts. 172-Pos.
6. Yang, Y., T. Sakamoto, Q. Xu, K. Thirumurugan, P. J. Knight, J. R. Sellers, (2005)
Characterization of Drosophila myosin VIIB. 2005 Biophysical Society Meeting
Abstracts. Biophysical Journal, Supplement, 647a, Abstract, 3177-Pos.
7. Thirumurugan, K., S.A. Burgess, J. Trinick, F. Wang, J.R. Sellers, H.D. White, P.J.
Knight, (2003) Temperature effects on myosin V head conformation. 2003
Biophysical Society Meeting Abstracts. Biophysical Journal, Supplement, 116a,
Abstract, 560-Pos.
8. Houmeida, A., B. Thompson, S.A. Burgess, J. Keen, K. Thirumurugan,
L.Tskhovrebova, P.J. Knight, J. Trinick, (2003) Preparation of synthetic titin endfilaments. 2003 Biophysical Society Meeting Abstracts. Biophysical Journal,
Supplement, 563a, Abstract, 2754-Pos.
Honors/Awards

Contribution Pay award from University of Leeds, UK

Wellcome Trust (UK) funded Post-doctoral fellowship

BBSRC (UK) funded Post-doctoral fellowship
28

CSIR Fellowship for Ph.D

BWX Ponniah Medal & Award for best Ph.D Thesis

Qualified National Eligibility Test (NET) conducted by CSIR

ASPEE fellowship for M.Sc
Membership
 Biophysical Society, USA (No. 24561)
 British Society for Cell Biology (No. 2724)
Invited Lectures
 Participated and gave a short talk at the Young Investigators Meeting (YIM2010) at
FFort, Raichak, Kolkata (February 8-12, 2010)
 Gave an invited talk at 'Molecular motors, Tracks and Transport' organized by
IMSc, NCBS (January 23-28, 2010)
Place & Date: VIT University, Vellore. August 19, 2010
Signature of the applicant
Kavitha Thirumurugan
29