Липкин Валерий Михайлович
Доктор химических наук, Профессор, Чл.-корр. РАН
Эл. почта: firstname.lastname@example.org
Работет в Институте биоорганической химии им. М. М. Шемякина и Ю. А. Овчинникова РАН с 1965 г. (аспирант, м. н. с., с. н. с. (1965—1986); заведующий лаборатории белков гормональной регуляции (1986—н. вр.); заместитель директора Института (1987—2008); руководитель Филиала ИБХ в г. Пущино (1987—2004).
Занимается педагогической деятельностью в Пущинском государственном университете, является деканом УЦ физико-химической биологии и биотехнологии; председатель Государственной экзаменационной комиссии на кафедре физико-химической биологии и биотехнологии МФТИ. Общее число публикаций — 240.
|Период обучения||Страна, город||Учебное заведение||Дополнительная информация|
|1959–1964||Россия, Москва||Московский химико-технологический институт им. Д.И. Менделеева (МХТИ)||Диплом инженера-технолога (химия)|
|1982||Россия, Москва||Присуждена учёная степень доктора химических наук|
Ведущий специалист в области химии белков и пептидов. Занимается исследованием белков, включенных в процессы трансмембранной передачи сигналов, дифференцировки и апоптоза клеток.
Премии и заслуги
- 1975: Премия Ленинского комсомола;
- 1981: Орден Дружбы народов;
- 1982: Государственная премия СССР в области науки и техники;
- 1997: Премия РАН имени Ю. А. Овчинникова.
Основные научные результаты
Липкиным В.М. и соавторами (под руководством академика Ю. А. Овчинникова) была установлена первая в нашей стране аминокислотная последовательность белка — аспартат-аминотрансферазы. В 1975—1985 гг. В. М. Липкин занимался исследованиями ДНК-зависимой РНК-полимеразы E. coli. Им с коллегами были определены аминокислотные последовательности всех субъединиц фермента, разработаны методы химической и фотохимической аффинной модификации его активных центров, выяснен молекулярный механизм ингибирования РНК-полимеразы антибиотиком рифампицином. Данные исследования внесли существенный вклад в выяснение механизмов процесса транскрипции.
Членство в научных обществах
- Член редакционной коллегии журнала «Биоорганическая химия»;
- член Научного совета РАН по биоорганической химии.
- (2017). Studying the Specific Activity of the Amide Form of HLDF-6 Peptide using the Transgenic Model of Alzheimer's Disease. Acta Naturae 9 (3), 64–70 [+]
The neuroprotective and nootropic activities of the amide form (AF) of the HLDF-6 peptide (TGENHR-NH2) were studied in transgenic mice of the B6C3-Tg(APPswe,PSEN1de9)85Dbo (Tg+) line (the animal model of familial Alzheimer's disease (AD)). The study was performed in 4 mouse groups: group 1 (study group): Tg+ mice intranasally injected with the peptide at a dose of 250 μg/kg; group 2 (active control): Tg+ mice intranasally injected with normal saline; group 3 (control 1): Tg- mice; and group 4 (control 2): C57Bl/6 mice. The cognitive functions were evaluated using three tests: the novel object recognition test, the conditioned passive avoidance task, and the Morris water maze. The results testify to the fact that the pharmaceutical substance (PhS) based on the AF of HLDF-6 peptide at a dose of 250 μg/kg administered intranasally efficiently restores the disturbed cognitive functions in transgenic mice. These results are fully consistent with the data obtained in animal models of Alzheimer's disease induced by the injection of the beta-amyloid (βA) fragment 25-35 into the giant-cell nucleus basalis of Meynert or by co-injection of the βA fragment 25-35 and ibotenic acid into the hippocampus, and the model of ischemia stroke (chronic bilateral occlusion of carotids, 2VO). According to the overall results, PhS based on AF HLDF-6 was chosen as an object for further investigation; the dose of 250 μg/kg was used as an effective therapeutic dose. Intranasal administration was the route for delivery.ID:1984
- (2016). Comparative study of the neuroprotective and nootropic activities of the carboxylate and amide forms of the HLDF-6 peptide in animal models of Alzheimer's disease. J. Psychopharmacol. (Oxford) 30 (1), 78–92 [+]
A comparative study of the neuroprotective and nootropic activities of two pharmaceutical substances, the HLDF-6 peptide (HLDF-6-OH) and its amide form (HLDF-6-NH2), was conducted. The study was performed in male rats using two models of a neurodegenerative disorder. Cognitive deficit in rats was induced by injection of the beta-amyloid fragment 25-35 (βA 25-35) into the giant-cell nucleus basalis of Meynert or by coinjection of βA 25-35 and ibotenic acid into the hippocampus. To evaluate cognitive functions in animals, three tests were used: the novel object recognition test, the conditioned passive avoidance task and the Morris maze. Comparative analysis of the data demonstrated that the neuroprotective activity of HLDF-6-NH2, evaluated by improvement of cognitive functions in animals, surpassed that of the native HLDF-6-OH peptide. The greater cognitive/ behavioral effects can be attributed to improved kinetic properties of the amide form of the peptide, such as the character of biodegradation and the half-life time. The effects of HLDF-6-NH2 are comparable to, or exceed, those of the reference compounds. Importantly, HLDF-6-NH2 exerts its effects at much lower doses than the reference compounds.ID:1387
- (2014). Human leukemia differentiation factor (HLDF) controls the cytokine-producing function of blood cells in gastric adenocarcinomas. Dokl. Biol. Sci. 454, 72–4 ID:1388
- (2013). Cloning and sequence analysis of cDNA encoding a new short form of rat acid chitinase. Dokl. Biochem. Biophys. 452 (1), 241–4 ID:1052
- (2012). Distinctive H-(RLDL)(4)-OH peptide complexes potentiate nanostructure self-assembling in water. Dokl. Biochem. Biophys. 443, 96–9 ID:949
- (2012). [Amyloid-like fibrils forming and fibroblasts destruction in Tenon's capsule in progressive myopia as a result of pigment epithelium derived factor resistance to restricted proteolysis]. Bioorg. Khim. 38 (6), 683–90 [+]
We have shown previously the presence of full length (50 kD) and truncated proteolytic form (45 kD) of pigment epithelium derived factor (PEDF) in the eye Tenon's capsule in progressive myopia. The full length PEDF is prevalent in myopia that correlates with breach in collagen fibrils forming. Immunohistochemical analysis of Tenon's capsule with polyclonal antibodies to PEDF revealed PEDF in control group being exclusively inside fibroblasts, whereas in myopia, PEDF was distributed extracellularly as halo around blasted fibroblasts. By means of atomic force microscopy and immunodot analysis with anti amyloid fibrils antibodies the ability was studied of recombinant PEDF fragments to form fibrils. Only full length PEDF was shown to form amyloid like fibril structures, but not the truncated form. Accumulation offibrils results in fibroblasts destruction and might be the cause of changes in biochemical and morphological structure of Tenon's capsule observed in myopia.ID:1053
- (2011). [Classification of self-organizing peptides]. Bioorg. Khim. 37 (6), 780–5 [+]
Amino acid sequences of known natural and synthetic self-assembling peptides were searched and analyzed for their characteristic patterns. The attempted formal numerical description of the repeating motifs, which have been revealed, resulted in building of general classification system embracing core-sequences of the peptides capable of nanostructure formation. Advantages and potency of the proposed rational classification were demonstrated via its comparison with the output from the earlier system described by the others.ID:950
- (2011). Novel protein haponin regulates cellular response to oxidative stress. Dokl. Biochem. Biophys. 440, 225–7 ID:1054
- (2009). Postischemic administration of HLDF-6 peptide ameliorates cognitive dysfunction and brain damage induced by chronic cerebral ischemia in rats. Dokl. Biol. Sci. 428, 418–22 ID:1056
- (2005). Secretion of the mammalian Sec14p-like phosphoinositide-binding p45 protein. FEBS J. 272 (21), 5595–605 [+]
Protein-lipid interactions are important for protein targeting, signal transduction, lipid transport, and the maintenance of cellular compartments and membranes. Specific lipid-binding protein domains, such as PH, FYVE, PX, PHD, C2 and SEC14 homology domains, mediate interactions between proteins and specific phospholipids. We recently cloned a 45-kDa protein from rat olfactory epithelium, which is homologous to the yeast Sec14p phosphatidylinositol (PtdIns) transfer protein and we report here that this protein binds to PtdIns(3,4,5)P3 and far weaker to less phosphorylated derivatives of PtdIns. Expression of the p45 protein in COS-1 cells resulted in accumulation of the protein in secretory vesicles and in the extracellular space. The secreted material contained PtdIns(3,4,5)P3. Our findings are the first report of a Sec14p-like protein involved in transport out of a cell and, to the best of our knowledge, inositol-containing phospholipids have not previously been detected in the extracellular space. Our findings suggest that p45 and phosphoinositides may participate in the formation of the protective mucus on nasal epithelium.ID:1055
- (2005). HLDF-6 peptide affects behavioral reactions and organism functions dependent on androgen hormones in normal and castrated male mice. Regul. Pept. 127 (1-3), 111–21 [+]
The hexapeptide Thr-Gly-Glu-Asn-His-Arg (HLDF-6), which was first identified as an active fragment of the human leukemia differentiation factor (HLDF) molecule, displays differentiation-inducing, neuroprotective and anti-drug abuse activities. Most of its in vivo effects were revealed only on male animals. We have studied HLDF-6 effects on a variety of organism functions and behavioral reactions, which are known to be dependent on androgen steroid hormones, both on castrated and normal (sham-operated) animals. Male NMRI mice were castrated or sham-operated at the age of 55 days (after puberty). After that, HLDF-6 peptide was injected daily during 3 weeks, followed by behavioral, morphological and biochemical testing. HLDF-6 increased testosterone level (1.5- to 2-fold) both in sham-operated and castrated animals. Sexual activity and pain sensitivity, which are strongly reduced in castrates, were completely or partially recovered by HLDF-6. At the same time, the peptide caused some effects similar to castration in sham-operated animals: aggression and locomotor activity were decreased; oral grooming was prolonged. Morphological studies of accessory sex glands showed that HLDF-6 partially normalizes the morphology and functional activity of seminal vesicles in castrates, but it does not prevent castration-induced apoptosis of prostate epithelial cells. Based on these observations, we can assume that HLDF-6 peptide displays at least two effects on androgen hormones metabolism in males: it stimulates testosterone biosynthesis by both testes and adrenals and simultaneously inhibits its conversion to dihydrotestosterone (DHT), most probably by diminution of 5alpha-reductase isoform 1 mRNA expression.ID:51
- (1999). A novel 45 kDa secretory protein from rat olfactory epithelium: primary structure and localisation. FEBS Lett. 450 (1-2), 126–30 [+]
cDNA clones encoding the 45 kDa protein were isolated from a rat olfactory epithelium cDNA library and their inserts were sequenced. The reconstructed protein sequence comprises 400 amino acids with a calculated molecular mass of 46,026 Da. A homology was revealed between the amino acid sequence of the 45 kDa protein and the proteins involved in the transfer of hydrophobic ligands. Using in situ hybridisation, the 45 kDa protein mRNA expression was detected in the layer of supportive cells of olfactory epithelium, apical region of trachea, surface layer of the ciliated bronchial epithelium in lung and in skin epidermis.ID:50
- (1994). A new human leukemia cell 8.2 kDa differentiation factor: isolation and primary structure. FEBS Lett. 356 (2-3), 327–9 [+]
A new 8.2 kDa differentiation factor has been purified to homogeneity from the cultural media of human myelogenous HL-60 leukemia cells induced by retinoic acid. cDNA clones encoding this factor were isolated from a cDNA library prepared from HL-60 differentiated cells and their nucleotide sequence has been determined. The deduced amino acid sequence of the differentiation factor molecule consists of 54 amino acid residues. The protein is shown to be glycosylated. It was shown by Northern blot experiments that the level of poly(A)+ RNA with a length of 450 nucleotides was higher in differentiated cells than in non-differentiated cells.ID:49
- (1993). The human rod photoreceptor cGMP phosphodiesterase beta-subunit. Structural studies of its cDNA and gene. FEBS Lett. 327 (3), 275–8 [+]
cDNA clones encoding the beta-subunit of the photoreceptor cGMP phosphodiesterase (PDE) were isolated from a human retina library and their sequence was determined. The encoded polypeptide consists of 854 amino acid residues with a calculated molecular mass of 98,416 Da. Alignment of the deduced amino acid sequence with the earlier analysed alpha-, beta- and alpha'-subunits of bovine and mouse PDEs demonstrates a high homology. Two overlapping recombinant lambda phage clones containing 26 kb of the human PDE beta-subunit gene were isolated from the genomic library. A total nucleotide sequence of exons 4-22 of the PDE beta-subunit gene was established which completely corresponded to the cDNA structure. According to sequence analysis no potential possibility for alternative splicing of the beta-subunit gene was observed between exons 20 and 21 which led to the formation of the beta'-subunit as described for mouse PDE. Polymerase chain reaction (PCR) experiments also confirm the absence of the PDE beta'-subunit in human retina.ID:48
- (1990). Beta-subunit of bovine rod photoreceptor cGMP phosphodiesterase. Comparison with the phosphodiesterase family. J. Biol. Chem. 265 (22), 12955–9 [+]
A group of cDNA clones encoding the beta-subunit of bovine rod photoreceptor cGMP phosphodiesterase were isolated for structural analysis. The encoded polypeptide has 853 residues with a calculated molecular mass of 98 kDa. The beta-subunit is 72% identical to the rod cGMP phosphodiesterase alpha-subunit. Like the alpha-subunit and the cone alpha'-subunit, the beta-subunit belongs to the family of phosphodiesterase genes. The beta- and alpha-subunits are more similar to each other than either is to the cone alpha'-subunit, suggesting either that the beta- and alpha-subunits diverged more recently or that their divergence was restrained by the rod functional environment.ID:47
- (1985). Complete amino acid sequence of gamma-subunit of the GTP-binding protein from cattle retina. FEBS Lett. 179 (1), 107–10 [+]
The complete amino acid sequence of the gamma-subunit of the GTP-binding protein from cattle retina has been established. The polypeptide chain of the gamma-subunit consists of 69 amino acid residues and contains the unusual sequence Cys35-Cys36. The Mr of the gamma-subunit is 8008.7.ID:55
- (1982). The primary structure of E. coli RNA polymerase, Nucleotide sequence of the rpoC gene and amino acid sequence of the beta'-subunit. Nucleic Acids Res. 10 (13), 4035–44 [+]
The primary structure of the E. coli rpoC gene (5321 base pairs) coding the beta'-subunit of RNA polymerase as well as its adjacent segment have been determined. The structure analysis of the peptides obtained by cleavage of the protein with cyanogen bromide and trypsin has confirmed the amino acid sequence of the beta'-subunit deduced from the nucleotide sequence analysis. The beta'-subunit of E. coli RNA polymerase contains 1407 amino acid residues. Its translation is initiated by codon GUG and terminated by codon TAA. It has been detected that the sequence following the terminating codon is strikingly homologous to known sequences of rho-independent terminators.ID:54
- (1977). ДНК-зависимая РНК-полимераза E.coli. Полная аминокислотная последовательность альфа-субъединицы. Биоорг. хим. 3 (1), 283–286 ID:147
- (1977). Primary structure of alpha-subunit of DNA-dependent RNA polymerase from Escherichia coli. FEBS Lett. 76 (1), 108–11 [+]
Transcription of genetic information in bacterial cells is mediated by DNA-dependent RNA polymerase (ribonucleoside-triphosphate:RNA nucleotidyltransferase, EC 126.96.36.199) [ 11. The enzyme from E. coliID:53
(mol. wt 500 000) has been shown to possess a complex structure  consisting of two large subunits, /?
and 0’ (mol. wt 155 000 and 16.5 000, respectively), two a-subunits (mol. wt 40 000) and initiation factor,
u (mol. wt 90 000). Its structural complexity is paralleled by the multistep nature of the transcription
process. Only limited information on the role of the individual subunits in the functioning of RNA polymerase
is available, owing in part to our lack of knowledge of its primary and spatial structure. We have, therefore,
undertaken an investigation into the primary structure of DNA-dependent RNA polymerase from
E. coli B. In the study reported here we have determined the complete amino acid sequence of the RNA poly
merase a-subunit (see  ) using a variety of degradation methods and i4C-enriched amino acid residues
for facilitating detection and isolation of the fragments. The polypeptide chain of the o-subunit has mol. w
36 512 and consists of 329 amino acid residues. In a comparative study of the peptide compositions of
various bacterial polymerase a-subunits it has been shown by peptide mapping that they are evolutionarily
conservative proteins .
- (1973). The complete amino acid sequence of cytoplasmic aspartate aminotransferase from pig heart. FEBS Lett. 29 (1), 31–34 [+]
Aspartate aminotransferase (L-aspartate: 2-oxo-glutarate aminotransferase, EC 188.8.131.52) is one of the principal pyridoxal-P-containing enzymes that catalyse the transamination reactions  representing key stepsID:52
at the intersection between the metabolic pathways of amino acids and dicarboxylic acids.
Although the catalytic mechanism of aspartate aminotransferase has been investigated at the level of substrate-coenzyme models , its elucidation in detail requires knowledge of the enzyme’s structure, considering, in particular, that the very high rates of the enzymic process are determined by the structural peculiarities of the specific protein(apoenzyme) of the aspartate aminotransferase. Accordingly, we embarked
on the task of elucidating the amino acid sequence of this enzyme. In the present paper the concluding stage
of the work is reported*. The object chosen for study was the aspartate aminotransferase
of the cytosol of pig heart; the enzyme, which is different from the mitochondrial isozyme
[5,6] was prepared by a previously reported procedure . The enzyme is a complex dimeric protein of
high molecular weight; each of the associated subunits consists of a single polypeptide chain and has no disulfide bridges. Indirect evidence (amino acid composition, analysis of N-terminal residues, and peptide maps) testified to the identity of the two subunits .
- (1966). Mass spectrometric determination of the amino-acid sequence of peptides. Nature 211 (5047), 361–6 [+]
Разработан новый метод определения аминокислотной последовательности в полипептидах с помощью масс-спектров эфиров их ацильных производных. Метод основан на анализе аминокислотного типа фрагментации пептидов с последовательным отщеплением остатков аминокислот и локализацией положительного заряда на фрагментах, несущих N-ацильную группу. Выяснены границы применимости метода и особенности, вносимые отдельными аминокислотными остатками в суммарную картину масс-спектра.ID:167