Dear Amyloid/Ageing subscribers:
I began to study Alzheimer's amyloid beta (Ab) protein in 1992, at
the time when it was becoming clear that this protein exists normally in a
soluble form (soluble Ab) and that it is not just a pathological protein. My
research activities yielded an on-going project, devoted to understanding in
more detail the normal biology of amyloid beta. Understanding the normal
biology of Ab would answer the questions of why soluble Ab does not undergo
fibrillogenesis in biological fluids but does polymerize into amyloid
fibrils in the disease, and what the biological consequences of Ab
deposition within the brain tissue are. This, in turn, would be crucial for
understanding the pathophysiology of Alzheimer's disease and for delineating
pathologically grounded new approaches to therapy.
My current postdoctoral position at NYU Medical Center just
expired. Thus, to continue my research I have to find another position. I
would be happy to continue my research as a part of your Department/
University/ Center, if you might wish to consider such a possibility.
Sincerely Yours,
Alexei R. Koudinov, M.D., Ph.D.
P.S. Enclosed please find CV and ABSTRACTS of my recent papers
______________________________________________________________________________
NAME: ALEXEI R. KOUDINOV
E.MAIL: koudin at imb.imb.ac.ru
EDUCATION:
University Years Degree Major
Moscow Medical University 1984- `MD Biochemistry,
1992
Cell/Molecular Biology, Medicine
Engelhardt Institute of 1992- PhD Biochemistry
Molecular Biology, Russian 1995
Academy of Sciences, Moscow.
( The Ph.D. research project was performed in the Department of Pathology,
New York University Medical Center. Ph.D. thesis: "Soluble amyloid beta is
complexed to high density lipoprotein 3 and very high density lipoproteins",
Sponsor/advisor: Professor B. Frangione, M.D., Ph.D.).
WORK EXPERIENCE:
1989 Laboratory Assistant, Genetics Department,
Moscow Medical University.
1990 Research Assistant, Enzymology Braunstein Memorial
Laboratory,
Engelhardt Institute of Molecular Biology.
1991-1992 Medical Assistant, Second Department of Neurology,
Moscow Medical University.
1992-present Assistant Research Scientist, Department of Pathology,
New York University Medical Center
RESEARCH EXPERIENCE:
1. Studies of structure-function relationship of two subforms of eukaryotic
Topoisomerase I (Masters' thesis research project, 1990-1992). Enzymology
Braunstein Memorial Laboratory, Engelhardt Institute of Molecular Biology,
Moscow, Russia. Sponsor: Dr. A. Gabibov, Ph.D.
Major techniques used: affinity, ion-exchange and hydrophobic interaction
chromatography, routinely and HPLC modification; protein quantification;
protein and DNA electrophoresis; isoelectrofocusing; enzymatic kinetic
studies; partial proteolysis; immunoblot analysis and enzyme-linked
immunosorbent analysis.
2. Studies of DNA-hydrolysing catalytic antibodies from autoimmune sera
(1991-1992) Enzymology Braunstein Memorial Laboratory, Engelhardt Institute
of Molecular Biology, Moscow, Russia. Advisors: Dr. A. Gabibov, Ph.D.
Major techniques used: IgG preparation from autoimmune sera by
chromatography, routine and HPLC modification; DNA isolation; DNA
hydrolysing activity assay.
3. Studies of autoimmune reaction in psoriatic skin (collaborative project
with Dr. A. Tkachenko, M.D., Ph.D., 1991-1992) Laboratory of cancerogenesis,
Engelhardt Institute of Moleculr Biology, Moscow, Russia.
Major techiques used: skin biopsy preparations; IgG preparation from human
plasma ( routine chromatography and HPLC on ProteinG Sepharose ); gel
electrophoresis and immunoblot analysis; 2D-gel electrophoresis.
4. Studies of connectivity of visual cortex in rats and human cortical
brain slices in vitro (Summer studentship program, Summer, 1992) Department
of Neurobiology, Brain Research Center, Weizmann Institute of Science,
Rehovot, Israel. Advisor: Dr. G. Kenan-Vaknin, Ph.D.
Major techniques used: preparation and maintaing in vitro mice, rat and
human cortical brain slices; probing of cortical connectivity in live brain
slices with tracer biocytin (a derivative of biotin); potential recording;
immunohistochemistry.
5. Studies of killing of Trypanosoma brucei by human serum (collaborative
project with Dr. S. Tomlinson, Ph.D., 1994-1995) Michael Heidelberger
Division of Immunology, Department of Pathology, New York University Medical
center, New York. Advisor: Dr. V. Nussenzweig, M.D., Ph.D.
Major techniques used: gradient ultracentrifugation; size exclusion
chromatography, FPLC modification; Trypanosoma brucei killing assay; gel
electrophoresis and immunoblot analysis.
6. Studies of normal biochemistry of soluble amyloid beta protein (Ph.D.
thesis research project, 1992-1996) Department of Pathology, New York
University Medical center, New York. Sponsor: Dr. B. Frangione, M.D., Ph.D.
Major techniques used: sequential flotation ultracentrifugation;
immunoaffinity chromatography; size exclusion and reverse phase HPLC;
thin-layer chromatography; gel electrophoresis and immunoblot analysis;
enzyme-linked immunosorbent analysis; chemiluminescence; optical and
chemiluminescent densitometry; amino acid sequence analysis; electron and
immunoelectron microscopy; immunocytochemistry; biotin technology for
tracing proteins in biological fluids; animal cell culture; metabolic
labelling of cell protein and lipid; lipid extraction; lyposome preparation;
enzymatic quantification of triglycerides, phospholipids, nonesterified and
total cholesterol; statistical analysis.
PUBLICATIONS:
KUDINOV, Alexei R., Bronstein, Igor B., Gabibov, Alexander G. and
Gololobov, Gennady V. (1992) Two subforms of eukaryotic topoisomerase I:
Purification and structure-function relationship. FEBS Lett 314: 267-270.
Tkachenko, Andrei V., Starkov, Ivan V., Shuster, Alexander M., KOUDINOV,
Alexei R., Zakharov, Sergei F., Shishkin, Sergei S., Mordovtsev, Vladimir
N., and Kisselev, Lev, L. (1992) Autoimmune reaction in psoriatic skin. In
Proceedings of the conference "Modern Enzymology: problems and trends".
St.Petersburg.
Ghiso, Jorge, Matsubara, Etsuro, KOUDINOV, Alexei, ChoiMiura, Nam Ho,
Tomita, Motono, Wisniewski, Thomas, Frangione, Blas. (1993). The
cerebrospinal fluid soluble form of Alzheimer's amyloid beta is complexed to
SP-40,40 (apolipoprotein J), an inhibitor of the complement membrane-attack
complex. Biochem J 293: 2730.
KOUDINOV, Alexei, Matsubara, Etsuro, Frangione, Blas, Ghiso, Jorge.
(1994). The Soluble Form of Alzheimer's Amyloid Beta Protein is Complexed to
High Density Lipoprotein 3 and Very High Density Lipoprotein in Normal Human
Plasma Biochem Biophys Res Commun 205: 1164-1171.
Tomlinson, Stephen, Jansen, Ana-Maria, KOUDINOV, Alexei, Ghiso, Jorge,
Choi-Miura, Nam-ho, Rifkin, Mary, Ohtaki, Sachiya, and Nussenzweig, Victor.
(1995) High-density-lipoprotein-independent killing of Trypanosoma brucei
by human serum. Molecular and Biochemical Parasitology 70: 131-138.
KOUDINOV, Alexei, R., Koudinova, Natalia, V., and Berezov, Temirbolat, T.
(1996). Alzheimer's peptides Ab1-40 and Ab1-28 inhibit the plasma
cholesterol esterification rate. Biochem Mol Biol Internat. 38(4): 747-752.
KOUDINOV, Alexei, R., Berezov, Temirbolat, T., and Koudinova, Natalia, V.
Multiple inhibitory effects of Alzheimer's peptide Ab1-40 on lipid
byosynthesis in cultured human HepG2 cells. In press. (SEE ABSTRACT BELOW)
KOUDINOV, Alexei, R., Koudinova, Natalia, V., Kumar, Asok, Beavis, Ronald,
and Ghiso, Jorge. (1996) Biochemical Biophysical Research Communication.
Biochemical characterization of Alzheimer's soluble amyloid beta protein in
human cerebrospinal fluid: association with high density lipoproteins.
Biochem Biophys Res Commun. In Press. (SEE ABSTRACT BELOW)
MEETING PRESENTATIONS:
KOUDINOV, Alexei, R., Koudinova, Natalia, V., Kumar, Asok. Is amyloid beta
an apolipoprotein? New York University Medical Center Neuroscience Meeting.
New York. December 1995.
KOUDINOV, Alexei, R., Koudinova, Natalia, V., Kumar, Asok, Beavis, Ronald,
and Ghiso, Jorge. Alzheimer's soluble amyloid beta protein is associated
with high density lipoproteins in normal human cerebrospinal fluid and is
secreted by HepG2 cells as a part of lipoprotein complexes. Society for
Neuroscience Annual Meeting. Washington, DC, 1996. Submitted. (SEE ABSTRACT
BELOW).
AWARDS:
1989-1992 All Russian Government Scholarship for outstanding success in
studies.
1992 Honorary Diploma of Medical Doctor. Moscow Medical University.
1992 Summer student fellowship. Weizmann Institute of Science. Rehovot. Israel
1994-1995 Pilot Research Grant from The Aging and Dementia Research Center
at NYU Medical Center. Project title: "HepG2 as a model for studies of sAb
secretion by human cells" (NIH Grant AG08051, Dr. S.Ferris, P.I.).
REFERENCES available upon request
10 June 1996
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MULTIPLE INHIBITORY EFFECTS OF ALZHEIMER'S PEPTIDE Ab1-40 ON
LIPID BIOSYNTHESIS IN CULTURED HUMAN HEPG2 CELLS.
Alexei R KOUDINOV(1), Temirbolat T BEREZOV(2) and Natalia V KOUDINOVA2,3*
Department of Pathology(1) and Medicine(3), New York University Medical Center,
560 First Avenue, TH427, New York, NY 10016, USA and Department of
Biochemistry(2), Russian Peoples' Friendship University School of Medicine,
Miklucho-Maklay St., 8, Moscow 117198, Russia
____________________________________________________________________________
BIOCHEMICAL CHARACTERIZATION OF ALZHEIMER'S SOLUBLE AMYLOID BETA
PROTEIN IN HUMAN CEREBROSPINAL FLUID: ASSOCIATION WITH
HIGH DENSITY LIPOPROTEINS.
Alexei R. Koudinov, Natalia V. Koudinova(1), Asok Kumar, Ronald C. Beavis(2)
and Jorge Ghiso
Departments of Pathology, Medicine(1) and Pharmacology(2), New York University
Medical Center, 560 First Avenue, New York, NY 10016
ABSTRACT
The soluble form of Alzheimer's amyloid b protein (sAb) is associated with
high density
lipoprotein (HDL) in normal human plasma (BBRC (1994) 205, 1164-71). Since
sAb is also
present in cerebrospinal fluid (CSF) and the lipoprotein pattern of CSF is
different from that of
plasma, it was of interest to ascertain whether the interaction of sAb with
HDL also occurs in
CSF. Normal human CSF lipoproteins were obtained by sequential flotation
ultracentrifugation
and analysed for the presence of sAb via immunoblot, size-exclusion
chromatography,
immunoelectron microscopy, N-terminal sequence and mass-spectrometry
analyses. Soluble
Ab was associated with CSF-HDL particles of 16.8 +/- 3.2 nm in diameter and
approximately 200
kDa of relative molecular mass. A ~4.3 kDa component purified by HPLC was
immunoreactive
with anti-Ab antibodies, exhibited an N-terminal sequence identical to the
Ab peptide and a
mass of 4325.1 Da, indicating that the main sAb specie associated with
CSF-HDL is sAb1-40.
_________________________________________________________________________
ALZHEIMER'S SOLUBLE AMYLOID BETA PROTEIN IS
ASSOCIATED WITH HIGH DENSITY LIPOPROTEINS IN NORMAL
HUMAN CEREBROSPINAL FLUID AND IS SECRETED BY HEPG2
CELLS AS A PART OF LIPOPROTEIN COMPLEXES
A. R. Koudinov*, N. V. Koudinova, A. Kumar, R. C. Beavis and J. Ghiso, New York
University Medical Center, TH 427, 560 First Avenue, New York, NY 10016.
ABSTRACT
The soluble form of amyloid b protein (sAb) is associated with high density
lipoprotein (HDL) in normal human plasma (Biochem Biophys Res Commun
(1994) 205, 1164-71). This suggests that sAb to HDL association is rather a
more general phenomenon taking place in other biological fluids and tissues.
To ascertain this hypothesis the colocalization of sAb with lipoproteins (LP)
was investigated in cerebrospinal fluid (CSF) and in cell culture supernatant.
Normal human CSF LPs were obtained by sequential flotation
ultracentrifugation and analyzed for the presence of sAb via immunoblot,
size-exclusion HPLC, immunoelectron microscopy, N-terminal sequence and
mass-spectrometry analyses. Soluble Ab was associated with ~200 kDa CSF-
HDL particles of 16.8 +/- 3.2 nm in diameter. A ~4.3 kDa component purified
by reverse phase HPLC was immunoreactive with anti-Ab antibodies,
exhibited an N-terminal sequence identical to the Ab peptide and a mass of
4325.1 Da, and therefore indicates that the main sAb specie associated with
CSF-HDL is sAb1-40. The sAb secretion by cells in association with LPs was
tested in human hepatoma HepG2 cell line. Soluble Ab in the cell culture
supernatant was detected in ~200 kDa molecular mass LP complexes in
association with apoJ, apoA-I, phospholipids, triglycerides and free and
esterified cholesterol. Our results suggest that the association of sAb with LP
represents a common mechanism for the peptide transport in biological
fluids.
_____________________________________________________________________________
Alexei Rudolphovich Koudinov
Institute of Molecular Biology, Russian Academy of Sciences
koudin at imb.imb.ac.ru
