This document is a resume for Dr. John Hsieh summarizing his background and experience. It lists his contact information, education, including a Ph.D. in Biochemistry from Washington University in St. Louis. It then details his professional experience as Laboratory Director at Icagen where he established facilities, invented an improved XRF assay apparatus, and developed assays for difficult to study ion channels. Prior to that, as Senior Scientist at Caldera Pharmaceuticals, he managed a team developing XRF technology, completed multiple SBIR grants, and developed assays for kinase activity and in vivo experiments. The resume emphasizes his expertise in enzymology, biochemistry techniques, and assay development.

1. JOHN
HSIEH,
PH.D.
2
Midland
Avenue
|
Franklin,
Massachusetts
02038
(505)
500-­‐7663
|
changtaihsieh@gmail.com
(https://www.linkedin.com/in/hsiehj)
SCIENTIST/SENIOR
SCIENTIST,
ENZYMOLOGY
A
self-­‐motivated,
independent,
and
innovative
biochemist
with
hands-­‐on
experience
with
mechanistic
enzymology,
elucidation
of
lead
molecule
mechanism
of
action,
and
assay
design.
Strong
communication,
problem-­‐solving,
and
analytical
skills.
Extensive
experience
working
with
multi-­‐disciplinary
teams.
Experience
in
scientific
operations
management,
mentoring,
and
project
management.
l Kinetic
mechanism
determination
l Allosteric
activation
or
inhibition
l Kd
and
Ki
determination
l Drug-­‐target
residence
times
l Kd
of
tight
binding
compounds
l Irreversible
enzyme
inactivation
ENZYMOLOGY
EXPERTISE
Steady-­‐state
kinetics
and
kinetic
equation
derivation,
transient
kinetic
methods
(stopped-­‐flow
and
chemical
quench-­‐flow)
and
kinetic
mechanism
determination
(KINSIM/FITSIM,
and
KinTek
Explorer),
mode
of
action
analysis,
kinetic
isotope
effects,
structure-­‐activity
analysis,
kinetic
assay
development,
ligand-­‐macromolecular
interaction
(Kd),
dissociation
kinetics
determination.
ADDITIONAL
TECHNICAL
EXPERTISE
l Biochemistry:
Protein
expression
and
purification,
molecular
biology,
liquid
chromatography
(HPLC,
FPLC,
IXC,
affinity,
size-­‐exclusion),
SDS-­‐PAGE,
protein
and
nucleic
acid
chemistry,
enzyme
immunoassays.
l Biophysical
Techniques:
Binding
thermodynamics,
Kd
determination
for
protein-­‐protein
and
compound-­‐
protein
interaction,
fluorescence
techniques
(time-­‐resolved
Fluorescence
Resonance
Energy
Transfer
(tr-­‐
FRET),
FRET,
polarization,
anisotropy,
competition),
SPR,
Dynamic
Light
Scattering.
l Assay
Development:
compound
screening
enzyme
activity
assays
by
UV/Vis,
radiological,
fluorescence,
and
fluorescence
anisotropy
techniques,
mid-­‐
and
high-­‐throughput
assays.
l Cell
Biology:
Mammalian
cell
cultures,
transfection,
cell-­‐based
ion
channel
and
transporter
assays.
l Computer
Skills:
GraphPad
Prism,
KinSim/FitSim,
KinTek
Explorer,
Microsoft
Office
Suite,
PyMOL.
l Other:
Mentoring,
grant
writing,
project
management,
scientific
operations.
EDUCATION
UNIVERSITY
OF
MICHIGAN,
Ann
Arbor,
Michigan,
Postdoctoral
Fellow
Advisor:
Carol
A.
Fierke;
Kinetic
Mechanism
and
Conformational
Dynamics
of
Ribonuclease
P
WASHINGTON
UNIVERSITY
SCHOOL
OF
MEDICINE,
St.
Louis,
Missouri,
Ph.D.,
Biochemistry
Doctoral
Thesis
Advisor:
Timothy
M.
Lohman;
Kinetic
Mechanisms
of
DNA
binding,
DNA
unwinding,
and
DNA-­‐stimulated
ATPase
Activity
by
the
Escherichia
coli
Rep
helicase
TEXAS
A&M
UNIVERSITY,
College
Station,
Texas,
B.Sc.
with
Honors,
Biochemistry
Undergraduate
Research
Advisor:
Paul
F.
Fitzpatrick;
Catalytic
Mechanism
of
Alcohol
Oxidase
EXPERIENCE
ICAGEN,
Inc.
(Formerly
XRpro
Sciences.)
Cambridge,
Massachusetts
Laboratory
Director,
Site
Manager
June
2013
to
December
2015
Worked
collaboratively
with
a
multi-­‐disciplinary
team
to
develop
proprietary
XRpro®
x-­‐ray
fluorescence
(XRF)
technology
as
a
label-­‐free
ion
channel
assay
for
small
molecule
ion
channel
modulator
screening.
My
invention
of
a

2. John
Hsieh,
Page
2
505-­‐500-­‐7663
384-­‐well
format
apparatus
enabled
Icagen
to
successfully
launch
XRpro®
high-­‐throughput
screening
services
in
2015.
• Established
Icagen’s
Cambridge
facility
and
managed
day-­‐to-­‐day
scientific
operations.
Oversaw
all
hiring
and
coordinated
contract
services.
Acquired
safety
permits
and
ensured
compliance.
• Invented
and
continued
to
improve
a
new
384-­‐well
XRF
assay
apparatus,
which
improved
XRF
sensitivity
by
>
100x
and
increased
throughput
by
20x.
• Developed
XRpro®
assays
for
ion
channels
and
transporters
targets
that
are
difficult
to
assay
using
standard
techniques.
Successfully
executed
8
projects
with
major
pharmaceutical
companies.
• Validated
XRpro®
assays
using
Transient
Receptor
Potential
ion
channels
(TRPA1,
TRPV1,
and
TRPC5).
Obtained
IC50
values
of
inhibitors
in
buffers
and
in
100%
serum
that
match
values
cited
in
the
literature.
• Validated
XRF
label-­‐free
chloride
channel
assay
using
GABAA
receptor
and
obtained
literature
EC50
value.
CALDERA
PHARMACEUTICALS.
Los
Alamos,
New
Mexico
Senior
Scientist
August
2009-­‐June
2013
Built
a
diverse
team
of
scientists
to
develop
x-­‐ray
fluorescence
XRpro®
technology
as
a
label-­‐free
bioinorganic
and
cell
biology
technique.
Completed
11
Small
Business
Innovation
Research
(SBIR)
projects
(>
$4.2M)
in
4
years.
• Supervise
scientific
operations
and
a
team
of
3
Ph.D.
scientists
and
1
technician.
• Grant
application
and
reports
preparation
(3
to
4
grants
per
year).
• Established
and
managed
a
peptide
sequencing
facility
with
$30,000
to
$40,000
annual
revenue.
• Developed
a
kinase
activity
assay
based
on
XRpro®
technology
for
a
pharmaceutical
company.
• Managed
successful
animal
testing
experiments
with
a
CRO
partner
for
a
$3M
NIAID
SBIR
grant
.
• Developed
XRF
based
novel
kinase
and
protease
substrate
discovery
from
a
pool
of
~
1
million
random
peptides.
This
work
led
to
a
service
that
Icagen
currently
offers.
• Represented
the
company
at
conferences
and
meetings
to
expand
the
business.
UNIVERSITY
OF
MICHIGAN
Ann
Arbor,
Michigan
Postdoctoral
Researcher,
Advisor:
Carol
A.
Fierke.
May
2002-­‐August
2009
Ribonuclease
P
(RNase
P)
is
an
essential
ribonucleoprotein
complex
that
catalyzes
the
maturation
of
tRNA's
from
precursors
(pre-­‐tRNA).
Used
binding
thermodynamics,
transient
kinetics,
nucleic
acid
chemistry,
and
trFRET
techniques
to
study
RNase
P’s
high
catalytic
efficiency
and
substrate
specificity.
• Initiated
fluorescence
assays
to
study
binding
thermodynamics
and
enzyme
kinetics
of
RNase
P.
• Determined
sub-­‐nanomolar
RNase
P-­‐pre-­‐tRNA
affinity
by
fluorescence
titration
and
Morrison
equation.
• Solved
the
kinetic
mechanisms
of
the
bacterial
and
the
yeast
nuclear
RNase
P’s
and
showed
similarities
and
differences
between
them.
• Determined
dissociation
kinetics
of
the
RNase
P-­‐pre-­‐tRNA
complex
by
kinetic
assays.
• Mentored
4
students
and
acted
as
a
consultant
in
enzymology
and
biophysical
studies
to
other
laboratories.
• Lectured
in
an
advanced
level
enzymology
class
(Chem
673,
in
2008
and
2009):
enzyme
inhibition,
motor
proteins,
and
ribozymes.
WASHINGTON
UNIVERSITY
SCHOOL
OF
MEDICINE,
St.
Louis,
Missouri
Ph.D.
Candidate,
Biochemistry,
Advisor:
Timothy
M.
Lohman.
August
1993-­‐May
2002
Combined
binding
thermodynamics,
transient
kinetics,
and
global
kinetic
data
analysis
to
study
ATPase
activity
and
DNA
unwinding
mechanisms
of
the
E.
coli
Rep
helicase.
• Solved
a
two-­‐site
ATPase
mechanism
of
the
Rep
helicase
homo-­‐dimer
by
global
kinetic
data
analysis
approach
(KINSIM/FITSIM).
Showed
that
ATP
hydrolysis
in
one
subunit
allosterically
activates
the
other
subunit.
• Obtained
the
first
high-­‐resolution
structure
of
Rep
DNA
helicase
complexed
with
single
stranded
DNA
and
ADP
(PDB
ID:
1UAA,
in
collaboration
with
Dr.
Gabriel
Waksman).
Gained
important
insights
to
helicase
structure,
domain
dynamics,
and
the
roles
of
conserved
“helicase
motifs”.

3. John
Hsieh,
Page
3
505-­‐500-­‐7663
• Analyzed
the
conserved
residues
at
the
ATPase
and
DNA
binding
sites
of
the
Rep
helicase,
and
identified
key
amino
acids
involved
in
ATP
binding
and
hydrolysis,
translocation,
and
unwinding
activity.
U.S.
PATENT
Peterson,
LJ.,
Mortillaro,
C.,
Warner,
W.,
Zahler,
N.
,
Hsieh,
J.,
Solomon,
E.,
and
Bharti,
P.
Method
for
Analysis
Using
X-­‐
ray
Fluorescence,
2015,
US
Patent
Number
9,063,066.
SELECTED
PUBLICATIONS
Wands,
A.M.,
Wang,
N.,
Lum,
J.
K.,
Hsieh,
J.,
Fierke,
C.
A.
&
Mapp,
A.
K.
(2011).
Transient-­‐state
kinetic
analysis
of
transcriptional
activator-­‐DNA
complexes
interacting
with
key
coactivator.
J
Biol
Chem
286,
16238-­‐45.
Hsieh,
J.,
Koutmou,
K.
S.,
Rueda,
D.,
Koutmos,
M.,
Walter,
N.
G.
&
Fierke,
C.
A.
(2010).
Divalent
cation
stabilizes
active
conformation
of
B.
subtilis
RNase
P
•
pre-­‐tRNA
complex:
the
role
for
inner-­‐sphere
metal
ion
in
RNase
P.
J
Mol
Biol
400,
38-­‐51.
Hsieh,
J.,
and
Fierke,
C.
A.
(2009).
Conformational
change
in
Bacillus
subtilis
RNase
P
holoenzyme•pre-­‐tRNA
complex
enhances
substrate
affinity
and
limits
cleavage
rate.
RNA
15,
1565-­‐77.
Hsieh,
J.,
Walker,
S.,
Fierke,
C.
A.,
and
Engelke,
D.
R.
(2008).
Pre-­‐tRNA
cleavage
by
yeast
nuclear
RNase
P
holoenzyme
is
rate-­‐limited
by
slow
product
release.
RNA
15,
224-­‐34.
Niranjanakumari,
S.,
Day-­‐Storms,
J.
J.,
Ahmed,
M.,
Hsieh,
J.,
Zahler,
N.
H.,
Venters,
R.
A.,
and
Fierke,
C.
A.
(2007)
Probing
architecture
of
B.
subtilis
RNase
P
holoenzyme
active
site
by
crosslinking
and
affinity
cleavage.
RNA
13:
521-­‐35.
Xiao,
S.,
Hsieh,
J.,
Nugent,
R.
L.,
Coughlin,
D.
J.,
Fierke,
C.
A.,
and
Engelke,
D.
R.
(2006)
Functional
characterization
of
conserved
amino
acids
in
Pop1p,
largest
common
protein
subunit
of
yeast
RNases
P
and
MRP.
RNA
12:
1023-­‐37.
Rueda,
D.,
Hsieh,
J.,
Day-­‐Storms,
J.
J.,
Fierke,
C.
A.,
and
Walter,
N.
G.
(2005)
5'
Leader
of
Precursor
tRNAAsp
Bound
to
Bacillus
subtilis
RNase
P
holoenzyme
has
extended
conformation.
Biochemistry
44:
16130-­‐9.
Cheng,
W,
Hsieh,
J,
Brendza,
K.M.,
and
Lohman,
T.M.
(2001)
E.
coli
Rep
oligomers
are
required
to
initiate
DNA
unwinding
in
vitro.
J
Mol
Biol.
310:327-­‐50.
Hsieh,
J.,
Moore,
K.J.M,
and
Lohman,
T.M.
(1999)
A
two-­‐site
kinetic
mechanism
for
ATP
binding
and
hydrolysis
by
E.
coli
Rep
helicase
dimer
bound
to
single-­‐stranded
oligodeoxynucleotide.
J
Mol
Biol
288:
255-­‐74.
Korolev,
S.,
Hsieh,
J.,
Gauss,
G.H.,
Lohman,
T.M.,
and
Waksman,
G.
(1997)
Major
domain
swiveling
revealed
by
crystal
structures
of
complexes
of
E.
coli
Rep
helicase
bound
to
single-­‐stranded
DNA
and
ADP.
Cell
90:
635-­‐47.
Wong,
I.,
Moore,
K.J.,
Bjornson,
K.P.,
Hsieh,
J.,
and
Lohman,
T.M.
(1996)
ATPase
activity
of
Escherichia
coli
Rep
helicase
dramatically
dependent
on
DNA
ligation
and
protein
oligomeric
states.
Biochemistry.
35:
5726-­‐34.
Menon,
V.,
Hsieh,
C-­‐T,
and
Fitzpatrick,
P.F.
(1995)
Substituted
Alcohols
as
Mechanistic
Probes
of
Alcohol
Oxidase.
Bioorganic
Chemistry
23:
42-­‐53.
Selected
Reviews
Hsieh
J,
Andrews,
A.J.,and
Fierke,
C.A.
(2004)
Roles
of
protein
subunits
in
RNA-­‐protein
complexes:
lessons
from
ribonuclease
P.
Biopolymers.
73:79-­‐89.
Lohman
T.M.,
Hsieh
J.,
Maluf
N.K.,
Cheng
W.,
Lucius
A.L.,
Fischer
C.J.,
Brendza
K.M.,
Korolev,
S.,
and
Waksman,
G.
(2004)
“DNA
Helicases,
Motors
that
Move
Along
Nucleic
Acids:
Lessons
from
SF1
Helicase
Superfamily”,
In
THE
ENZYMES,
3rd
ed.,
Volume
XXIII,
Energy
Coupling
and
Molecular
Motors,
edited
by
Hackney,
D.D.
and
Tamanoi,
F.,
pp.
304-­‐364.
AFFILIATIONS
RNA
Society,
American
Chemical
Society,
Biophysical
Society