ABSTRACT
The fraction of proteins capable of binding to photoreceptor membranes in a Ca2+-dependent manner was isolated from bovine rod outer segments. One of these proteins with apparent molecular mass of 32 kD (p32) was purified to homogeneity and identified as annexin IV (endonexin) by MALDI-TOF mass-spectrometry. In immunoblot, annexin IV purified from bovine rod outer segments cross-reacted with antibodies against annexin IV from bovine liver. This is the first detection of annexin IV in vertebrate retina.
Subject(s)
Annexin A4/analysis , Retinal Rod Photoreceptor Cells/chemistry , Amino Acid Sequence , Animals , Annexin A4/chemistry , Annexin A4/immunology , Annexin A4/isolation & purification , Cattle , Cross Reactions/immunology , Immunoblotting , Liver/chemistry , Molecular Sequence Data , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Annexins are phospholipid-binding proteins and are abundant in the lung. Annexins I and IV, but not II and VI, have been detected in bronchoalveolar lavage (BAL) fluids from calves inoculated with Pasteurella haemolytica, the pathogen for calf pneumonia. In this study, BAL fluids from calves with experimental pneumonia induced by inoculation to right lung lobes of bovine herpes virus-1 (BHV-1), the major viral pathogen for pneumonia, were examined for detection of annexins I and IV. Of 6 calves inoculated with BHV-1, annexins I and IV were coincidentally detected in BAL fluids from right lung lobes of 4 calves, but not in BAL fluids from left lung lobes of 6 inoculated calves or those from left and right lung lobes of 3 control calves. Annexin II and VI were not found in any BAL fluids examined. These results, together with previous findings on calves inoculated with Pasteurella haemolytica, suggest that the release of annexins I and IV onto the alveolar surface is an essential event occurring in response to pulmonary infections of BHIV-1 and Pasteurella haemolytica.
Subject(s)
Annexin A1/isolation & purification , Annexin A4/isolation & purification , Bronchoalveolar Lavage Fluid/chemistry , Cattle Diseases/metabolism , Herpesviridae Infections/veterinary , Herpesvirus 1, Bovine/chemistry , Pneumonia, Viral/veterinary , Animals , Annexin A1/blood , Blotting, Western/veterinary , Bronchoalveolar Lavage/veterinary , Cattle , Cattle Diseases/virology , Chromatography, DEAE-Cellulose/veterinary , Chromatography, Gel/veterinary , Electrophoresis, Polyacrylamide Gel/veterinary , Herpesviridae Infections/metabolism , Lung/chemistry , Male , Pneumonia, Viral/metabolismABSTRACT
We have previously demonstrated that annexin IV, one of the calcium/phospholipid-binding annexin family proteins, binds to glycosaminoglycans (GAGs) in a calcium-dependent manner (Kojima, K., Yamamoto, K., Irimura, T., Osawa, T., Ogawa, H., and Matsumoto, I. (1996) J. Biol. Chem. 271, 7679-7685). In this study, we investigated the GAG binding specificities of annexins IV, V, and VI by affinity chromatography and solid phase assays. Annexin IV was found to bind in a calcium-dependent manner to all the GAG columns tested. Annexin V bound to heparin and heparan sulfate columns but not to chondroitin sulfate columns. Annexin VI was adsorbed to heparin and heparan sulfate columns in a calcium-independent manner, and to chondroitin sulfate columns in a calcium-dependent manner. An N-terminal half fragment (A6NH) and a C-terminal half fragment (A6CH) of annexin VI, each containing four units, were prepared by digestion with V8 protease and examined for GAG binding activities. A6NH bound to heparin in the presence of calcium but not to chondroitin sulfate C, whereas A6CH bound to heparin calcium-independently and to chondroitin sulfate C calcium-dependently. The results showed that annexin IV, V, and VI have different GAG binding properties. Some annexins have been reported to be detected not only in the cytoplasm but also on the cell surface or in extracellular components. The findings suggest that the some annexins function as recognition elements for GAGs in extracellular space.
Subject(s)
Annexin A4/metabolism , Annexin A5/metabolism , Annexin A6/metabolism , Glycosaminoglycans/metabolism , Amino Acid Sequence , Animals , Annexin A4/chemistry , Annexin A4/isolation & purification , Annexin A5/chemistry , Annexin A5/isolation & purification , Annexin A6/chemistry , Annexin A6/isolation & purification , Brain/metabolism , Calcium/pharmacology , Cattle , Chondroitin Sulfates/metabolism , Chromatography, Affinity , Heparin/metabolism , Heparitin Sulfate/metabolism , Humans , Liver/metabolism , Molecular Sequence Data , Sequence Alignment , Sequence Homology, Amino Acid , Substrate SpecificityABSTRACT
The structure of a trigonal crystal form of N-terminally truncated [des-(1-9)] bovine annexin IV, an annexin variant that exhibits the distinctive property of binding both phospholipids and carbohydrates in a Ca2+-dependent manner, has been determined at 3 A (0.3 nm) resolution -space group: R3; cell parameters: a=b=118.560 (8) A and c=82.233 (6) A-. The overall structure of annexin IV, crystallized in the absence of Ca2+ ions, is highly homologous to that of the other known members of the annexin family. The trimeric assembly in the trigonal crystals of annexin IV is quite similar to that found previously in non-isomorphous crystals of human, chicken and rat annexin V and to the subunit arrangement in half of the hexamer of hydra annexin XII. Moreover, it resembles that found in two-dimensional crystals of human annexin V bound to phospholipid monolayers. The propensity of several annexins to generate similar trimeric arrays supports the hypothesis that trimeric complexes of such annexins, including annexin IV, may represent the functional units that interact with membranes.
Subject(s)
Annexin A4/chemistry , Amino Acid Sequence , Animals , Annexin A4/analogs & derivatives , Annexin A4/isolation & purification , Cattle , Crystallization , Crystallography, X-Ray , Hydrogen Bonding , Kidney/chemistry , Models, Molecular , Molecular Sequence Data , Phospholipids/metabolism , Protein Binding , Protein Conformation , Protein Structure, Secondary , Protein Structure, TertiaryABSTRACT
Se realizó una revisión de la s 245 pacientes enviadas a Colposcopia por citología PAP II más cambios virales, en un período de 3 años, se encontró en la biopsia dirigida un 68.16 por ciento de lesión por condiloma, un 31.42 por ciento asociado a NIC y un 0.4 por ciento con cáncer invasor. Esto confirma los informes de la relación de PVH y cáncer de cérvix y por lo tanto la necesidad de colposcopia y biopsia en las pacientes con citologías PAP II y cambios virales (coilocitos, disqueratocitos, binucleación)
