Matti Vuento - Academia.edu (original) (raw)
Papers by Matti Vuento
Biophysical Journal, 2004
Infection and Immunity, 1984
Purified cathepsin G fragments of fibronectin were used to locate the binding sites for streptoco... more Purified cathepsin G fragments of fibronectin were used to locate the binding sites for streptococci and staphylococci in the fibronectin molecule. The iodinated, NH2-terminal, 30-kilodalton (kd) fragment bound to group A and G streptococci and to Staphylococcus aureus. The 125I-labeled, COOH-terminal, 120- to 140-kd fragment bound weakly to group A streptococcus strain and to S. aureus when tested in a buffer of low ionic strength. The 30- and 120- to 140-kd fragments inhibited the binding of iodinated fragments to bacteria. The two fragments were, on a molar basis, equally effective, and they were more potent inhibitors than intact fibronectin. The gelatin-binding 40-kd fragment neither bound to any of the bacterial strains nor inhibited the binding of 125I-labeled 30-kd or 125I-labeled 120- to 140-kd fragments to bacteria. The results indicate that fibronectin has at least two separate binding sites for streptococci and staphylococci, one in the NH2-terminal region and another in...
Biochemical Society Transactions, 1981
The enzyme transferring the oligosaccharide from Dol-PP-GlcNAc2Man Glc3 to asparagine residues of... more The enzyme transferring the oligosaccharide from Dol-PP-GlcNAc2Man Glc3 to asparagine residues of glycoproteins has been solubilized from yeast membranes. Enzyme activity was tested by measurtransfer of the glycosyl moiety from Dol-PP-; " C ] saccharides to the hexapeptide Tyr-Asn-Leu-Thr-Ser-Val. The enzyme transfers besides dolichyldiphosphate-bound GlcNAc2Man Glc3 also GlcNAc Man, and GlcNAc2; the rate decreases in this order. No transfer is observed from Dol-PP-GlcNAc2Mang and from Dol-PP-GlcNAc. A clear difference in K values for the hexapeptide was observed, with $ifferent dolichol-linked sugar derivatives. With Dol-PP-GlcNAc2 a peptide concentration of 0.6 mM resulted in half maximal transfer rate, whereas 0.05 mM peptide were sufficient with Dol-PP-GlcNAc2Man Glc 9 9 2 as donor. 9 3
Protides of the Biological Fluids, 1987
ABSTRACT Electrofusion method has been used to produce monoclonal antibodies against polypeptide ... more ABSTRACT Electrofusion method has been used to produce monoclonal antibodies against polypeptide antigens. Suggestions of some optimum conditions for electrically-mediated splenocyte-myeloma hybridizations are presented.
Infection and Immunity, 1985
The attachment of Staphylococcus aureus (Cowan I) and two strains of group A and G streptococci o... more The attachment of Staphylococcus aureus (Cowan I) and two strains of group A and G streptococci on glass cover slips coated with fibronectin, fibronectin fragments, or fibrinogen was studied. The attachment was quantitated by counting the attached bacteria on glass surfaces coated with a similar molarity of the proteins. Fibronectin was a more effective attachment factor than fibrinogen for staphylococci, while group G streptococci attached better on fibrinogen- than on fibronectin-coated cover slips. In this system, group A streptococci bound almost exclusively to substrate-bound fibrinogen. Attachment experiments involving the use of staphylococci pretreated with soluble fibronectin or fibrinogen revealed that bacterium-bound fibronectin and fibrinogen were able to enhance the adherence on cover slips coated with fibronectin. The 30-kilodalton NH2-terminal and the 120- to 140-kilodalton COOH-terminal fragments of fibronectin, both of which contain bacterial binding sites, mediated...
FEBS Letters, Dec 14, 2004
A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories f... more A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories feline kidney (NLFK) cells of the capsid proteins of canine parvovirus (CPV), VP1 and VP2, or corresponding counterparts fused to EGFP. VP1 and VP2, when expressed alone, both had equal nuclear and cytoplasmic distribution. However, assembled form of VP2 had a predominantly cytoplasmic localization. When VP1 and VP2 were simultaneously present in cells, their nuclear localization increased. Thus, confocal immunofluorescence analysis of cells transduced with the different baculovirus constructs or combinations thereof in the absence or presence of infecting CPV revealed that the VP1 protein is a prerequisite for efficient targeting of VP2 to the nucleus. The baculovirus vectors were functional and the genes of interest efficiently introduced to this CPV susceptible mammalian cell line. Thus, we show evidence that the system could be utilized to study targeting of the CPV capsid proteins.
International Journal of Nanomedicine, 2010
Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, ... more Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, and oncolytic cancer virotherapy. Here we have studied cell death mechanisms of canine parvovirus (CPV) to increase the knowledge on the CPV life cycle in order to facilitate the development of better parvovirus vectors. Morphological studies of CPV-infected Norden laboratory feline kidney (NLFK) cells and canine fibroma cells (A72) displayed characteristic apoptotic events. Apoptosis was further confirmed by activation of caspases and cellular DNA damage. However, results from annexin V-propidium iodide (PI) labeling and membrane polarization assays indicated disruption of the plasma membrane uncommon to apoptosis. These results provide evidence that secondary necrosis followed apoptosis. In addition, two human cancer cell lines were found to be infected by CPV. This necrotic event over apoptotic cell death and infection in human cells provide insightful information when developing CPV as a nanotool for cancer treatments.
Biochemical Society Transactions, 1981
Glial-Neuronal Communication in Development and Regeneration, 1987
Outgrowth of neurites during development and regeneration in central and peripheral nervous syste... more Outgrowth of neurites during development and regeneration in central and peripheral nervous systems involves mechanisms based on soluble neurite-promoting molecules, like the nerve growth factor (1) and other diffusible substances (2–6). Adhesion of neurons to other cells or to the surrounding extracellular matrix is also thought to play an important role in the outgrowth of neurites (7). Thus, specialized regions of the neurons identified as growth cones adhere to surfaces of other cells or to other extracellular surfaces, and these adhesions are thought to guide axonal growth (7).
Virus Research, 2008
ISBN 978-951-39-5633-2 (nid.) ISBN 978-951-39-5634-9 (PDF) Yhteenveto: Virusproteiinit ja niiden ... more ISBN 978-951-39-5633-2 (nid.) ISBN 978-951-39-5634-9 (PDF) Yhteenveto: Virusproteiinit ja niiden vuorovaikutukset isäntäsolussa Diss. Viruses are ancient parasites that predate all three domains of life: Eukarya, Bacteria and Archaea. Viruses usually specifically infect a determined cell type, largely defined by the receptors they recognise. Canine parvovirus is a small, non-enveloped animal virus that infects cells in dividing cells, especially in puppies. PRD1 is a bacteriophage infecting a wide range of Gram-negative bacteria. It is a well-known model virus of the Tectiviridae family and its structure has been the subject of numerous studies. In this thesis, interactions between virus proteins and the host cell were studied using both CPV and PRD1 as model viruses. The study of the complex inner protein localization in bacterial cells is in its infancy, yet particularly important for the understanding of virus-cell interactions. We constructed a vector library for the production of fluorescent fusion proteins and used it to study the localization of several PRD1 viral proteins inside E. coli. Results revealed variations in the localization patterns; one dividing characteristic appeared to be the multimericity of proteins. The vector library, together with complementation assays, was further utilized in the study of PRD1 proteins P33 and P17. The obtained results indicated that these proteins interact with the host cell chaperonin machinery. The GroEL/GroES machinery is vital for both the host cell and several phages including PRD1. Finally, the interactions of the CPV capsid with lipid membranes were studied in order to elucidate the virus escape mechanism from the endocytic vesicles. Results suggested that, together with known phospholipase A2 activity, there is an additional, membrane-induced mechanism to promote the escape.
Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like par... more Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like particles
Correspondence: Matti Vuento Department of Biological and Environmental Science, Survontie 9, PO ... more Correspondence: Matti Vuento Department of Biological and Environmental Science, Survontie 9, PO Box 35, FIN – 40351 University of Jyväskylä, Finland Email matti.vuento@bytl.jyu.fi Abstract: A rapid method and instrumentation for quantifi cation of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The fi rst anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the fi rst anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane...
Biochemical and biophysical research communications, 2005
Although sharing a T=1 icosahedral symmetry with other members of the Parvoviridae family, it has... more Although sharing a T=1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence correlation spectroscopy showed that an average of nine EGFP domains were associated with these virus-like structures. Atomic force microscopy and immunoprecipitation studies showed that EGFP was displayed on the surface of these fVLPs. Confocal imaging indicated that these chimeric complexes were targ...
Biol Chem, 2002
Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major e... more Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major envelope glycoprotein gp64 of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) were characterized by fluorescence correlation spectroscopy (FCS). FCS detected Brownian motion of single, intact recombinant baculovirus display particles with a diffusion coefficient (D) of (2.89±0.74)10 8 cm2s 1 and an apparent hydrodynamic radius of 83.35±21.22 nm. In the presence of sodium dodecyl sulfate (SDS), Triton X-100, and octylglucoside, the diffusion time was reduced to the 0.2 ms range (D = 7.5710 7 cm2s 1), showing that the fusion proteins were anchored in the viral envelope. This allowed for a calculation of the number of single gp64 fusion proteins incorporated in the viral membrane. A mean value of 3.2 fluorescent proteins per virus particle was obtained. Our results show that FCS is the method of choice for studying enveloped viruses such as a display virus with one component being GFP.
Chemistry and Physics of Lipids, Feb 28, 2010
The effects of tri-and monoglycerides on phospholipid (POPC) membranes were studied using spectro... more The effects of tri-and monoglycerides on phospholipid (POPC) membranes were studied using spectroscopical methods. Triolein was found to form two types of POPC-rich membranes, both with POPC or as a three-component system with monopalmitin. These two membrane types were determined as co-existing phases based on their spontaneous and stable separation and named heavy and light phase according to their sedimentation behaviour. Marked differences were seen in the physical properties of these phases, even though only minor compositional variation was detected. The light, less polar phase was found to be less ordered and more fluid and seemed to allow significantly lower amount of water penetration into the membrane-water interface than pure POPC membrane. The heavy phase, apart from their slightly altered water penetration, resembled more a pure POPC membrane. As triglycerides are present in lysosomal membranes, the present results can be seen as an implication for polarity-based water permeability barrier possibly contributing to the integrity of lysosomes.
Biophysical Journal, 2004
Infection and Immunity, 1984
Purified cathepsin G fragments of fibronectin were used to locate the binding sites for streptoco... more Purified cathepsin G fragments of fibronectin were used to locate the binding sites for streptococci and staphylococci in the fibronectin molecule. The iodinated, NH2-terminal, 30-kilodalton (kd) fragment bound to group A and G streptococci and to Staphylococcus aureus. The 125I-labeled, COOH-terminal, 120- to 140-kd fragment bound weakly to group A streptococcus strain and to S. aureus when tested in a buffer of low ionic strength. The 30- and 120- to 140-kd fragments inhibited the binding of iodinated fragments to bacteria. The two fragments were, on a molar basis, equally effective, and they were more potent inhibitors than intact fibronectin. The gelatin-binding 40-kd fragment neither bound to any of the bacterial strains nor inhibited the binding of 125I-labeled 30-kd or 125I-labeled 120- to 140-kd fragments to bacteria. The results indicate that fibronectin has at least two separate binding sites for streptococci and staphylococci, one in the NH2-terminal region and another in...
Biochemical Society Transactions, 1981
The enzyme transferring the oligosaccharide from Dol-PP-GlcNAc2Man Glc3 to asparagine residues of... more The enzyme transferring the oligosaccharide from Dol-PP-GlcNAc2Man Glc3 to asparagine residues of glycoproteins has been solubilized from yeast membranes. Enzyme activity was tested by measurtransfer of the glycosyl moiety from Dol-PP-; " C ] saccharides to the hexapeptide Tyr-Asn-Leu-Thr-Ser-Val. The enzyme transfers besides dolichyldiphosphate-bound GlcNAc2Man Glc3 also GlcNAc Man, and GlcNAc2; the rate decreases in this order. No transfer is observed from Dol-PP-GlcNAc2Mang and from Dol-PP-GlcNAc. A clear difference in K values for the hexapeptide was observed, with $ifferent dolichol-linked sugar derivatives. With Dol-PP-GlcNAc2 a peptide concentration of 0.6 mM resulted in half maximal transfer rate, whereas 0.05 mM peptide were sufficient with Dol-PP-GlcNAc2Man Glc 9 9 2 as donor. 9 3
Protides of the Biological Fluids, 1987
ABSTRACT Electrofusion method has been used to produce monoclonal antibodies against polypeptide ... more ABSTRACT Electrofusion method has been used to produce monoclonal antibodies against polypeptide antigens. Suggestions of some optimum conditions for electrically-mediated splenocyte-myeloma hybridizations are presented.
Infection and Immunity, 1985
The attachment of Staphylococcus aureus (Cowan I) and two strains of group A and G streptococci o... more The attachment of Staphylococcus aureus (Cowan I) and two strains of group A and G streptococci on glass cover slips coated with fibronectin, fibronectin fragments, or fibrinogen was studied. The attachment was quantitated by counting the attached bacteria on glass surfaces coated with a similar molarity of the proteins. Fibronectin was a more effective attachment factor than fibrinogen for staphylococci, while group G streptococci attached better on fibrinogen- than on fibronectin-coated cover slips. In this system, group A streptococci bound almost exclusively to substrate-bound fibrinogen. Attachment experiments involving the use of staphylococci pretreated with soluble fibronectin or fibrinogen revealed that bacterium-bound fibronectin and fibrinogen were able to enhance the adherence on cover slips coated with fibronectin. The 30-kilodalton NH2-terminal and the 120- to 140-kilodalton COOH-terminal fragments of fibronectin, both of which contain bacterial binding sites, mediated...
FEBS Letters, Dec 14, 2004
A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories f... more A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories feline kidney (NLFK) cells of the capsid proteins of canine parvovirus (CPV), VP1 and VP2, or corresponding counterparts fused to EGFP. VP1 and VP2, when expressed alone, both had equal nuclear and cytoplasmic distribution. However, assembled form of VP2 had a predominantly cytoplasmic localization. When VP1 and VP2 were simultaneously present in cells, their nuclear localization increased. Thus, confocal immunofluorescence analysis of cells transduced with the different baculovirus constructs or combinations thereof in the absence or presence of infecting CPV revealed that the VP1 protein is a prerequisite for efficient targeting of VP2 to the nucleus. The baculovirus vectors were functional and the genes of interest efficiently introduced to this CPV susceptible mammalian cell line. Thus, we show evidence that the system could be utilized to study targeting of the CPV capsid proteins.
International Journal of Nanomedicine, 2010
Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, ... more Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, and oncolytic cancer virotherapy. Here we have studied cell death mechanisms of canine parvovirus (CPV) to increase the knowledge on the CPV life cycle in order to facilitate the development of better parvovirus vectors. Morphological studies of CPV-infected Norden laboratory feline kidney (NLFK) cells and canine fibroma cells (A72) displayed characteristic apoptotic events. Apoptosis was further confirmed by activation of caspases and cellular DNA damage. However, results from annexin V-propidium iodide (PI) labeling and membrane polarization assays indicated disruption of the plasma membrane uncommon to apoptosis. These results provide evidence that secondary necrosis followed apoptosis. In addition, two human cancer cell lines were found to be infected by CPV. This necrotic event over apoptotic cell death and infection in human cells provide insightful information when developing CPV as a nanotool for cancer treatments.
Biochemical Society Transactions, 1981
Glial-Neuronal Communication in Development and Regeneration, 1987
Outgrowth of neurites during development and regeneration in central and peripheral nervous syste... more Outgrowth of neurites during development and regeneration in central and peripheral nervous systems involves mechanisms based on soluble neurite-promoting molecules, like the nerve growth factor (1) and other diffusible substances (2–6). Adhesion of neurons to other cells or to the surrounding extracellular matrix is also thought to play an important role in the outgrowth of neurites (7). Thus, specialized regions of the neurons identified as growth cones adhere to surfaces of other cells or to other extracellular surfaces, and these adhesions are thought to guide axonal growth (7).
Virus Research, 2008
ISBN 978-951-39-5633-2 (nid.) ISBN 978-951-39-5634-9 (PDF) Yhteenveto: Virusproteiinit ja niiden ... more ISBN 978-951-39-5633-2 (nid.) ISBN 978-951-39-5634-9 (PDF) Yhteenveto: Virusproteiinit ja niiden vuorovaikutukset isäntäsolussa Diss. Viruses are ancient parasites that predate all three domains of life: Eukarya, Bacteria and Archaea. Viruses usually specifically infect a determined cell type, largely defined by the receptors they recognise. Canine parvovirus is a small, non-enveloped animal virus that infects cells in dividing cells, especially in puppies. PRD1 is a bacteriophage infecting a wide range of Gram-negative bacteria. It is a well-known model virus of the Tectiviridae family and its structure has been the subject of numerous studies. In this thesis, interactions between virus proteins and the host cell were studied using both CPV and PRD1 as model viruses. The study of the complex inner protein localization in bacterial cells is in its infancy, yet particularly important for the understanding of virus-cell interactions. We constructed a vector library for the production of fluorescent fusion proteins and used it to study the localization of several PRD1 viral proteins inside E. coli. Results revealed variations in the localization patterns; one dividing characteristic appeared to be the multimericity of proteins. The vector library, together with complementation assays, was further utilized in the study of PRD1 proteins P33 and P17. The obtained results indicated that these proteins interact with the host cell chaperonin machinery. The GroEL/GroES machinery is vital for both the host cell and several phages including PRD1. Finally, the interactions of the CPV capsid with lipid membranes were studied in order to elucidate the virus escape mechanism from the endocytic vesicles. Results suggested that, together with known phospholipase A2 activity, there is an additional, membrane-induced mechanism to promote the escape.
Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like par... more Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like particles
Correspondence: Matti Vuento Department of Biological and Environmental Science, Survontie 9, PO ... more Correspondence: Matti Vuento Department of Biological and Environmental Science, Survontie 9, PO Box 35, FIN – 40351 University of Jyväskylä, Finland Email matti.vuento@bytl.jyu.fi Abstract: A rapid method and instrumentation for quantifi cation of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The fi rst anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the fi rst anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane...
Biochemical and biophysical research communications, 2005
Although sharing a T=1 icosahedral symmetry with other members of the Parvoviridae family, it has... more Although sharing a T=1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence correlation spectroscopy showed that an average of nine EGFP domains were associated with these virus-like structures. Atomic force microscopy and immunoprecipitation studies showed that EGFP was displayed on the surface of these fVLPs. Confocal imaging indicated that these chimeric complexes were targ...
Biol Chem, 2002
Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major e... more Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major envelope glycoprotein gp64 of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) were characterized by fluorescence correlation spectroscopy (FCS). FCS detected Brownian motion of single, intact recombinant baculovirus display particles with a diffusion coefficient (D) of (2.89±0.74)10 8 cm2s 1 and an apparent hydrodynamic radius of 83.35±21.22 nm. In the presence of sodium dodecyl sulfate (SDS), Triton X-100, and octylglucoside, the diffusion time was reduced to the 0.2 ms range (D = 7.5710 7 cm2s 1), showing that the fusion proteins were anchored in the viral envelope. This allowed for a calculation of the number of single gp64 fusion proteins incorporated in the viral membrane. A mean value of 3.2 fluorescent proteins per virus particle was obtained. Our results show that FCS is the method of choice for studying enveloped viruses such as a display virus with one component being GFP.
Chemistry and Physics of Lipids, Feb 28, 2010
The effects of tri-and monoglycerides on phospholipid (POPC) membranes were studied using spectro... more The effects of tri-and monoglycerides on phospholipid (POPC) membranes were studied using spectroscopical methods. Triolein was found to form two types of POPC-rich membranes, both with POPC or as a three-component system with monopalmitin. These two membrane types were determined as co-existing phases based on their spontaneous and stable separation and named heavy and light phase according to their sedimentation behaviour. Marked differences were seen in the physical properties of these phases, even though only minor compositional variation was detected. The light, less polar phase was found to be less ordered and more fluid and seemed to allow significantly lower amount of water penetration into the membrane-water interface than pure POPC membrane. The heavy phase, apart from their slightly altered water penetration, resembled more a pure POPC membrane. As triglycerides are present in lysosomal membranes, the present results can be seen as an implication for polarity-based water permeability barrier possibly contributing to the integrity of lysosomes.