Curriculum Vitae
Name: Saroj Kumar
Sex: Male
Professional address: Laboratory for Structure and Function of Biological Membranes
Universite Libre de Bruxelles, Campus Plaine, Building BC, Boulevard
duTriomphe, 1050 Brussels, Belgium, +32-471******,
**********.*@*****.***
Career Objective
A challenging position requiring the innovation, hard work, consultancy and expertise
attained in field of science.
Education, main scientific appointments
Postdoctoral fellow (Dec.2011-present) Laboratory for Structure and Function of Biological
Membranes, Université Libre de Bruxelles, Brussels, Belgium
Histology of breast cancer improved by infrared imaging
Research assistant (Aug-Nov.2011), Department of Biochemistry and Biophysics,
Stockholm University, Sweden
PhD (2011) Department of Biochemistry and Biophysics, Stockholm University, Sweden,
Advisor: Professor Andreas Barth
Thesis: “Infrared spectroscopy: Method development and ligand binding to pyruvate
kinase”
Side projects:
Energy coupling in the Ca2+-ATPase (mainly expression and purification of
recombinant ATPase) and H+, K+-ATPase studied by Fourier transform IR
(FTIR) spectroscopy
Study of yeast and bacterial cells by attenuated total reflection (ATR)-FTIR
spectroscopy
Licentiate degree (2010) Department of Biochemistry and Biophysics, Stockholm
University, Sweden
Advisor: Professor Andreas Barth
Thesis: “Infrared spectroscopy: Method development and ligand binding studies”
Degree project (2006) Department of Biochemistry and Biophysics, Stockholm University,
Sweden
Redox linked conformational changes in cytochrome c and Protein-herbicide
interaction studied by ATR-FTIR spectroscopy
Indian council of medical research fellow (2004) Institute of pathology ICMR, Safdarjung,
New Delhi, India
Microscopic colitis studied by electron microscopy
Master of Science in Biophysics (2004) All India institute of medical sciences, New Delhi,
India
Extensive course work in theoretical and molecular biophysics
Master Thesis: “Cloning of cobra venom phospholipase A2”
Bachelor of Science in human biology (2001) All India institute of medical sciences, New
Delhi, India
anatomy, biochemistry, biophysics, pathology, microbiology, physical and
chemical sciences
Professional activities
20% teaching duty of PhD (since 2007) and supervision of graduate and postgraduate
students
Supervising high school students during summer training (since 2008)
Attended 5 workshops and 16 conferences on spectroscopy and structural proteomics
Collaboration with a large chemical company (2005): plant herbicide study by ATR-
FTIR
PhD and postdoctoral scholarships
Brains back to Brussels (2011) to pursue postdoc (only in exceptional cases given to
people not coming from Belgium)
Postdoctoral grant from National fund for scientific research, Belgium (2011)
Swedish postdoctoral grant (2011) to pursue postdoc studies
Marie Curie International outgoing fellowship for career development (2011),
proposal was placed on the reserve list
Lawski fellowship (2006-2009) to pursue PhD study
Other scholarships and awards
W. Bagge och Rhodin donationstipendium (2009): 1400 Euro project grant
K & A Wallenbergs stiftelse (2008) travel grant to participate in the 5th International
Conference on Advanced Vibrational Spectroscopy, Melbourne, Australia
C. F. Liljevalch J:ors (2008) travel grant to participate in 2nd international workshop on
expression, structure and function of membrane Proteins, Florence, Italy
Techniques worked with
Time resolved Fourier transform infrared spectroscopy (FTIR)
FTIR microspectroscopy
Attenuated total reflection FTIR spectroscopy
UV-visible spectroscopy
Circular dichroism spectroscopy
Fluorescence spectroscopy
X-Ray crystallography
Eukaryotic cell culture
Cell culture of human cell lines
Expression and purification of membrane proteins (Ca2+-ATPase)
Cloning, sequencing, polymerase chain reaction (PCR)
Electrophoresis, Western blot
Monolayer preparation of lipids
Electron microscopy
Crystallization
Histopathology slide preparation and microscopical studies
Computation skills
Application software related to processing of scientific data, MS office, Origin, Pymol, and
Kinemage, Matlab based program Kinetics
Research results
Publications/Manuscripts
S.Kumar, C.Desmedt, D.Larsimont, C.Sotiriou, E.Goormaghtigh: Change in the
microenvironment of breast cancer studied by FTIR imaging. Analyst, 2013, 138,
4058-65
S.Kumar, C.Li and A.Barth: Conformational changes of recombinant Ca2+-ATPase
studied by reaction-induced infrared difference spectroscopy. FEBS J. 2013, 13 Feb,
1-10
A.Valdivia, A.Barth, Y.Batista and S.Kumar: Characterization of recombinant
antibodies for cancer therapy by infrared spectroscopy. Biologicals, 2013, 1-7
S.Kumar, N.Eremina and A.Barth: Detection of Ligand Binding to Proteins through
Observation of Hydration Water. J. Phys. Chem. B. 2012, 116, 13968-74
S.Kumar and A.Barth: Allosteric effects of fructose bisphosphate on muscle pyruvate
kinase studied by infrared spectroscopy. J. Phys. Chem. B. 2011, 115 (39), 11501-05
S. Kumar and A. Barth: Effects of ions on ligand binding to pyruvate kinase: mapping
the binding site with infrared spectroscopy. J. Phys. Chem. B. 2011, 115, 6784-89
S.Kumar and A.Barth: Following enzyme activity with infrared spectroscopy. Sensors
2010, 10, 2626-37
S.Kumar and A.Barth: Phosphoenolpyruvate and Mg2+ Binding to Pyruvate Kinase
Monitored by Infrared Spectroscopy. Biophys. J., 2010, 98, 1931-40
M.Rudbeck, S.Kumar, M. Blomberg, M-A. Mroginski, A. Barth: Infrared spectrum of
phosphoenol pyruvate: computational and experimental studies. J. Phys. Chem. A.
2009, 113, 2935-42
M.Krasteva, S.Kumar and A.Barth: A dialysis accessory for attenuated total reflection
IR spectroscopy. Spectroscopy (cover page) 2006, 20, 89-94
T.S.Shabi, S.Kumar, E. Goormaghtigh, Y. Geerts: Role of solubilizing group removal
rate on the grain size and crystallinity of diketopyrrolopyrrole based compounds and
its impact on their optical and electrical properties. Crystal growth and design,
Accepted
C.Li, S.Kumar and A.Barth: Effect of E309Q on Ca2+-ATPase pump studied by
reaction-induced Infrared difference spectroscopy. Manuscript in preparation
S.Singh, S.Kumar, A.Barth and M. Malmsten: Membrane and lipopolysaccharide
interactions of C-terminal peptides from S1 peptidases by infrared spectroscopy.
Manuscript in preparation
Reviews/Books
S.Kumar: Infrared spectroscopy is a physical tool to study biomolecules. Applied
spectroscopy reviews, 49 (3), 2014, 187-200 (online)
S.Kumar: Infrared spectroscopy: Method development and ligand binding to pyruvate
kinase. 2011,, pp 1-57, ISBN: ***-**-****-297-4
Reviewer for scientific journals
Spectroscopy letters
Environmental science and technology
Molecular pharmaceutics
International Conferences/Workshops (International)
1. S.Kumar and E. Goormaghtigh (2012): Conference Spec 2012, “Shedding new light
on disease”. Chaing Mai, Thailand
2. S.Kumar, and A.Barth (2009): 2nd international workshop on expression, structure and
function of membrane proteins. Poster presentation and actively participate in
workshop. Florence, Italy
3. S.Kumar and A.Barth (2009): Infrared studies of ligand binding and enzyme activity
of biomoleules. Poster presentation at 5th International conference on advanced
vibrational spectroscopy, Melbourne, Australia
4. S.Kumar, and A.Barth (2008): Ligand induced conformational changes of pyruvate
kinase study by ATR-FTIR spectroscopy. Poster presentation at XXIX European
conference on molecular spectroscopy, Opatija, Croatia
5. S.Kumar (2008): Workshop in protein production for structural genomics, at Structural
genomics consortium (SGC), Stockholm
6. S.Kumar, and A.Barth (2007): Protein-ligand interaction study by ATR-FTIR
spectroscopy. Poster presentation at 12th European conference on the spectroscopy of
biological molecules, Paris, France
7. S.Kumar, M.Krasteva and A.Barth (2005): Protein-ligand interaction study by ATR-
FTIR spectroscopy. Poster presentation at 11th European conference on the
spectroscopy of biological molecules, Aschaffenburg, Germany
Oral presentations (External)
1. 2013: 15th European Conference on the Spectroscopy of Biological Molecules,
Oxford, UK
2. 2010: CEA, Saclay, Gif sur Yvette, France
3. 2010: 30th European Congress on Molecular Spectroscopy. Florence, Italy
Major accomplishments in research
Method development for infrared spectroscopy
I have taken part in successfully developing the three methods for infrared spectroscopy. The
first method couples a dialysis accessory to an attenuated total reflection unit to study ligand
binding to macromolecules. The main advantage of this method is the possibility to
manipulate the sample of interest during an experiment and to perform multiple experiments
with different ligands on the same sample which saves time and material. This work has been
published (#10). We developed a second method for infrared spectroscopy which provides a
direct, monitor of enzymatic reactions of three enzymes. The main advantage of the infrared
method is that it observes the reaction of interest directly, i.e. no activity assay is required that
converts the progress of the reaction into an observable quantity as often necessary for
spectroscopic detection in the UV/visible spectral range. This work has been published in
(#7). The third method is a novel approach to detect ligand binding by a change in water
absorption. The idea behind this work was that water is displaced in all ligand binding process
from the hydration shell of proteins and ligands to bulk water. Infrared spectroscopy has a
high sensitivity for water interactions and any changes in the hydrogen bond pattern can be
detected. Using the absorption of the OH stretching mode we were able to detect these
changes. This work has been published (#4).
The molecular mechanism of pyruvate kinase (PK)
PK is an important enzyme of the glycolytic pathway that catalyses the transfer of phosphate
from phosphoenolpyruvate (PEP) to adenosine diphosphate. By using infrared spectroscopy,
we observed that all types of secondary structures are affected upon PEP binding to PK. We
observed that the enzymatic environment affected the structure of the bound substrate, but
none of the bonds of PEP were significantly distorted upon binding, in particular not those of
the reacting phosphate group. We also studied the effects of monovalent and divalent cations
on binding of the substrate PEP to PK, as both types of cations are necessary for PK's activity.
We observed that the binding mode of PEP is affected more by different divalent cations than
by different monovalent cations, which shows that the divalent cation plays a more decisive
role for the structural consequences of PEP binding. Mn2+ showed very interesting effects as
PEP binding produced larger signals than with the physiological ion Mg2+, indicating a more
closed conformation after PEP binding. Furthermore, we studied the effects of the allosteric
effector fructose 1, 6 bisphosphate (FBP) on PEP binding to PK in presence of different ion
combinations. The results indicate that FBP binding shifts the equilibrium of the
PK:PEP:Mg2+:K+ complex towards the closed conformation. The PEP concentration for
saturating signal was lower with FBP than without which shows that the affinity of PK for
PEP increased by binding of FBP. This work has resulted in four publications (#5, 6, 8, 9). My
contribution was the planning and execution of the project.
Energy coupling in the Ca2+-ATPase
The Ca2+-ATPase is a P-type ATPase which pumps two calcium ions across the sarcoplasmic
reticulum membrane at the expense of the hydrolysis of one ATP molecule. The aim of the
project was to understand how chemical energy from ATP is converted to directional Ca2+
transport by the Ca2+-ATPase by studying the effects of amino acid substitutions. As an
essential first step, I established in our laboratory the expression and purification system of
recombinant Ca2+-ATPase and obtained active protein. It was characterized by biophysical
methods and compared with the native Ca2+-ATPase. This work represents the majority of my
work during my PhD project. It was done in the settings of a biophysics group without
previous experience in molecular biology and initially without appropriate equipment. This
work has resulted in one publication (#2) and a manuscript of it is in preparation. My
contribution was the independent establishing of the expression and purification in our
laboratory from a published protocol.
Histology of breast cancer improved by infrared imaging
Breast cancer is a global public health issue. At diagnosis, 90% of the patients appear to have
operable breast cancer, that is, the disease that is confined to the breast and ipsilateral axilla.
However, only up to 40% are cured by the operation while around 50% of these women will
develop metastatic disease (incurable). Although different tools have been developed to assist
clinicians in selecting patients who should receive adjuvant therapy, it still remains a
challenge to distinguish those patients who would really need adjuvant systemic therapy from
those who could be spared such a treatment. Fourier transform infrared (FTIR) imaging is a
suitable technique for the quantitative analysis of tissue in the clinical routine. This
quantitative analysis should allow a description of the most common cell types found on
histological slices and a grading of tumor tissues from microscopic observation of the slices,
presence of markers and clinical observations. Particular attention is paid to the influence of
the microenvironment (stroma) on cancer progression. I found a spectral marker which
distinguishes the infected and normal stroma which resulted a publication (#1).