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F Kanemitsu

Publications and source records attributed to F Kanemitsu.

At least 19 recordsLinked to original sources
PubMed1994

Characterization of human creatine kinase BB and MB isoforms by means of isoelectric focusing.

Isoforms of creatine kinase (creatine-N-phosphotransferase, CK, EC 2.7.3.2), BB and MB, were isolated from healthy human brain tissue and cardiac muscle, respectively, and were characterized by means of isoelectric focusing (IEF). CK-BB isoforms in Tris-HCl buffer were focused at pI 4.5 (a tissue form) and those in fresh sera from healthy adults were focused at pI 5.0, 5.1 and 5.2 (plasma forms). The IEF patterns of CK-BB isoforms were not altered following treatment with carboxypeptidase B and incubation in fresh serum at 37 degrees C; thus, it was found that there was no lysine at the C-terminal of the CK-B subunit and CK-BB isoforms were not results of the removal of lysine. Three CK-BB isoforms in fresh sera were identified to be oxidized, intermediate and reduced forms from the anodal side, respectively, by treatment with hydrogen peroxide and 2-mercaptoethanol. The oxidized form of CK-BB seemed to have higher affinity to IgG than the other two plasma forms of CK-BB. On the other hand, CK-MB isoforms in Tris-HCl buffer were focused at pI 5.4 (a tissue form) with a minor band at pI 5.2 and those in sera were focused at pI 5.0, 5.1, 5.2 (plasma forms) and 5.4. Four CK-MB isoforms were identified to be reduced, intermediate and oxidized forms without lysine from the anodal side, respectively, and the cathodal band was a tissue form with lysine.

Brain Chemistry↗
PubMed1992

Creatine kinase MB isoforms for early diagnosis and monitoring of acute myocardial infarction.

MB isoforms of creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2, CK) in 848 sera obtained from 80 patients with acute myocardial infarction (AMI) were studied by agarose gel isoelectric focusing. In 173 sera (20%) from 25 patients (31%), a new isoform designated as MB3 (pI 5.4) was detected at the cathodal side of MB2 (pI 5.2) in addition to the previously known MB2 and MB1 (pI 5.1). The new isoform MB3 was found in the extract of the cardiac muscle. MB3 was dominant in the sera at an earlier stage and a shorter period of time after AMI: 1-14 h (range), 1-29.5 h and 4-154 h for MB3, MB2 and MB1 dominant, respectively. MB3 was therefore found to be an earlier and a shorter phase indicator for AMI than MB2 or MB1. However, MB2 greater than 1 was the most prevalent pattern at the time of admission to the hospital. In AMI, specificity was 96.2%, 92.4% and 90.6%, and sensitivity was 20.4%, 88.9% and 97.6%, for MB3, MB2 and MB1 isoforms, respectively. CK-MB isoform patterns were biased to MB1 dominant in the deceased group, and to MB2 dominant in the surviving group. Therefore determination of CK-MB isoforms is also useful in the course of observation of AMI. The fourth isoform, MB0 (pI 5.0), was detected at the anodal side of MB1. MB0 was a minor band of the CK-MB isoform which appeared when serum CK-MB activity increased.

Acute Disease↗
PubMed1991

An immunologic pathway for intravascular catabolism of creatine kinase subform MM3.

In vitro incubation of the MM3 subform of human creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2, CK) with fresh human serum resulted in the formation of a complex of high relative molecular mass (Mr 320 kDa). The formed complex (macro CK-MM3) consists of both CK-MM3 and immunoglobulin A (IgA), and its amount of the formed complex was proportional to CK-MM3 activity and IgA concentration. Two molecules of CK-MM3 combined with one molecule of IgA, and the immunoglobulin inhibited the enzyme activity. As IgA does not form complexes with other subforms (CK-MM2 and CK-MM1) or CK-MB, the antigen specificity of IgA to CK-MM3 is definitely exacting. The circumstantial evidence suggests that macro CK-MM3 is a specific antigen-antibody complex. Macro CK-MM3 was detected in all of the examined sera of adult patients with more than 2001 U/1 CK activity (the positive percentage of macro CK-MM3 in all adult patients was 73%), but not detected in sera of patients who were younger than 12 months old. No relationship was observed between macro CK-MM3 and the patients' underlying diseases. Macro CK-MM3 formation suggested to be an immunologic pathway for intravascular catabolism of CK-MM3 when its activity increases.

Adolescent↗
PubMed1990

Clinical application of subforms of creatine kinase MM and macro creatine kinases.

The subforms of MM isozyme of creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2, CK) in sera obtained from healthy adults and patients were determined by agarose gel isoelectric focusing (IEF). The patients were classified into six groups according to serum CK-MM activities and IEF patterns. The IEF spectra offered useful information on cell hyperplasia, augmented cell membrane permeability, cell destruction and release time of CK-MM in the circulation from the cells for diagnosis, progress observation and prognosis, especially in the cases of chronic hepatic diseases, acute myocardial infarction and muscular dystrophy. Macro CKs were also determined by IEF. Macro CKs could be completely distinguished from each other, and CK isozymes consisting of macro CK type 1 could be presumed by isoelectric points.

Chromatography, Gel↗
PubMed1990

Electrophoretic and kinetic characterization of three variants of soluble cytoplasmic L-alanine:2-oxoglutarate aminotransferase in human liver tissue.

Three common variants of soluble cytoplasmic L-alanine:2-oxoglutarate aminotransferase (ALT, EC 2.6.1.2), sALT 1, 2-1 and 2, were isolated from normal human liver, and characterized by electrophoretic and kinetic analyses. The isoelectric point of sALT 1 was pH 6.45. sALT 2-1 was focused into three bands with pl 6.1, 6.2 and 6.45; sALT was focused into one band with pl 6.1. The electrophoretic mobilities of sALTs altered to the fast beta-globulin fraction after aging or papain treatment. Ammonia was produced during the latter, and the altered migration was considered to be caused by deamidation of sALT. The relative molecular mass of each of the enzymes was 110,000. Minor differences in the apparent Km values among the multiple forms for both L-alanine and 2-oxoglutarate were observed after incubation with 100 mumol/L of pyridoxal phosphate (PALP). PALP stimulation of the enzyme activities was also different. sALT 1 was more stable than sALT 2-1 and 2 after heat and urea treatments. In human sera from 1065 adult Japanese, sALT 2-1, a heterozygote form of sALT 1 and 2, was dominant.

Alanine Transaminase↗
PubMed1988

Creatine kinase isoenzymes.

Analytical methods for and the clinical significance of ATP:creatine-N-phosphotransferase (EC 2.7.3.2, CK) isoenzymes have been described. The main assay methods for the isoenzymes are electrophoresis, ion-exchange chromatography and immunoinhibition. Electrophoresis is a time-consuming technique, but it is essential for the identification of CK isoenzymes. Ion-exchange chromatography is reliable for the determination of CK-MB and useful for the isolation and purification of CK isoenzymes. The disadvantages of this method are overlapping of isoenzymes with neighbouring fractions and the inevitable dilution of enzyme activity by the elution buffer. Immunoinhibition is a rapid, sensitive, specific and quantitative technique. However, mitochondrial CK and CK-immunoglobulin complex remain after the inhibition of the M subunit with the antibodies. Thus, when assessing the experimental results, it is necessary to be certain that a false positive for CK-MB has not been obtained. Since each technique has both merits and demerits, the technique most suited to the particular laboratory should be selected.

Animals↗
PubMed1987

Isoenzyme specificities of immunoglobulins isolated from lactate dehydrogenase-immunoglobulin complexes.

Lactate dehydrogenase (L-lactate: NAD oxidoreductase, EC 1.1.1.27, LD) specific immunoglobulins were isolated from LD-immunoglobulin complexes in sera from 11 patients and the isoenzyme specificity of the immunoglobulins was studied by sandwich electroimmunofixation (SEIF). The immunoglobulins differed in isoenzyme specificity, depending on the heavy and/or light chains. IgA kappa recombined with LD2 and LD3, and was thought to be the antibody to common structures (H2M) in LD2 (H3M) and LD3 (H2M2). IgA kappa in a IgA/G kappa lambda case recombined with LD3-5, so that the IgA kappa might be an antibody to M2 dimer in the isoenzymes. IgA lambda recombined only with LD3, and was thought to recognize the H2M2 structure of LD3. As IgG kappa and/or lambda recombined with all LD isoenzymes of LD1-5, the immunoglobulins might react with common structures in H and M subunits. IgG lambda recombined with LD2-5, particularly with LD5, so that the IgG lambda was thought to be an antibody to the M subunit in LD2-5. From the above findings and the tetrameric structure of the LD molecule, 18 types of antibodies to different antigenic determinants on LD molecules can be theoretically derived. Each of the present 11 cases of immunoglobulin corresponded to one of the 18 types, respectively. A formula which can theoretically determine the percent LD activity recombined with immunoglobulins is described.

Antibody Specificity↗
PubMed1986

Clinical significance and characteristics of creatine kinase-immunoglobulin complexes in sera from patients with malignant tumors.

Creatine kinase (CK, ATP: creatine N-phosphotransferase, EC 2.7.3.2) and immunoglobulin complexes were detected in 27 of 234 (11.5%) patients with malignant tumors. The complexes electrophoresed between CK-MM and the sample application point. The positive rate was significantly higher than rates in patients with benign diseases, or in healthy adults (p less than 0.001). Primary malignancies of the 27 patients were colon 8, gastric 6, pancreatic 4, liver 5, pulmonary 3 and mammary 1. As 22 of the 27 patients had a recurrence or a metastasis and the mortality rate of the positive group 6 mth after the tests was high (85%), most of the 27 patients had advanced tumors. Thus, the CK-immunoglobulin complexes seem to be a prognosticator of patients with advanced tumors. The classes and types of immunoglobulins in the complexes were IgA lambda, 7; IgA kappa lambda, 5; IgA (types of the light chain were not identified), 4; IgA/G kappa, 1; IgA/G lambda, 3; and IgA/G kappa lambda, 7. The molecular sizes of the complexes were 384,000 +/- 22,000 (mean +/- 1 SD). The results of recombination assays using IgA isolated from the complexes of 4 patients suggested that the complexes in the patients' sera were mitochondrial CK-IgA. Non CK-M subunit activities of the positive group were 58 +/- 40 U/l (mean +/- 1 SD) and they were significantly higher than those of the complex negative group of tumor patients (p less than 0.01). This is probably the first report of detection of mitochondrial CK-IgA complexes.

Adult↗
PubMed1986

Sandwich electroimmunofixation (SEIF) for the assessment of isoenzyme specificities of immunoglobulins isolated from enzyme-immunoglobulin complexes.

Sandwich electroimmunofixation (SEIF) was used to determine immunologic specificity for isoenzymes of immunoglobulins isolated from enzyme-immunoglobulin complexes. After electrophoretic separation of isoenzymes, isoenzyme-specific human immunoglobulins obtained from complexes and antihuman immunoglobulin antibodies were applied to the supporting medium and allowed to react. Complexes of isoenzyme-immunoglobulins-anti-immunoglobulin antibodies formed immunoprecipitates and were fixed in the supporting medium. After washout of the unreacted enzymes and proteins with buffer, the immunoprecipitates were stained. A comparison with control enzymograms allowed for a determination as to whether the immunoglobulins reacted with specific isoenzymes. When an immunologic reaction with more than 2 isoenzymes occurred, the specificity was quantitated by densitometry. SEIF, used to examine immunoglobulins isolated from aspartate aminotransferase-, lactate dehydrogenase-, alkaline phosphatase- and amylase-immunoglobulin complexes, was found to be a rapid and reliable technique. This approach showed that immunoglobulins differ in their specificities for various isoenzymes.

Alkaline Phosphatase↗
PubMed1986

[CK anomaly].

Explore the source record for details and available documents.

Creatine Kinase↗
PubMed1985

Comparison of enzymatic characteristics of creatine kinase BB from human healthy and tumor stomach tissues.

Creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2, CK) BB isoenzyme from stomach tumor tissue was partially purified and its characteristics were compared with those from healthy tissue. Molecular mass of tumor CK-BB was estimated to be 82 000 by polyacrylamide gel electrophoresis. Tumor CK-BB was separated into 2 main subbands around pH 4.5 and 11, minor subbands around pH 5-7.5 by agarose isoelectric focusing. The isoenzyme reacted with anti-human brain CK-BB antibodies and formed a hybrid, CK-MB, with CK-MM prepared from healthy human skeletal muscle. The above physicochemical and immunological characteristics of tumor CK-BB were the same as those of normal CK-BB from normal stomach tissue. Optimum pH of tumor CK-BB was more acidic than that of normal CK-BB. Affinity for creatine phosphate and heat sensitivity of tumor CK-BB were slightly lower than those of normal CK-BB. Tumor CK-BB was more stable after iodoacetamide and urea treatments.

Brain↗
PubMed1984

Mitochondrial creatine kinase as a tumor-associated marker.

Sera from 167 patients with malignant tumor were examined with regard to mitochondrial creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2, CK). The CK (CKm) was detected in 66 cases (40%), in 42 of 96 men and in 24 of 71 women. Seventeen patients had hepatic carcinomas and 14 gastric carcinoma. CKm was also present in sera from patients with malignancy of the lung, the mammary gland etc. In the CKm positive group, about 85% had metastatic and/or infiltrating malignancy, and about 90% were undergoing chemotherapy. The figures resembled those in the CKm negative group. Mortality rates in the CKm positive group were 1.7 times higher than in the CKm negative group. CKm may be a marker of some biological factors leading to and/or resulting from a poor prognosis and not simply of primary or metastatic tumors. Most of the total serum CK activities in the CKm positive group were within normal limits. CKm activities in the gastric carcinoma group were higher than those in other groups, particularly in comparison to the liver carcinoma group.

Adult↗
PubMed1983

A new creatine kinase found in mitochondrial extracts from malignant liver tissue.

A new creatine kinase (CK, ATP: creatine N-phosphotransferase, EC 2.7.3.2) band was found in mitochondrial extracts prepared from malignant liver tissue. By agar gel electrophoresis the new band (CKmLT) migrated between CK-MM and a sample application point. The CK band was not found in mitochondrial extracts of normal liver, cytosolic extracts of normal and malignant liver homogenates. We thus presume that the origin of CKmLT is from mitochondria of malignant liver tumors. CKmLT reacted with anti-mitochondrial CK antibodies so that its antigenicity was similar to that of mitochondrial CK. Physico-chemical characteristics were similar to those of the polymeric form of the normal liver mitochondrial CK (CKmL1); i.e. relative molecular mass (320 000 to 350 000), heat stability, and the behavior in 2 mol/l urea. CKmLT migrated to the CKmL1 and the MM position after long-term storage at -20 degrees C. CKmLT did not demonstrate affinity to concanavalin A, and there was no significant change in the electrophoretic mobility of CKmLT following neuraminidase treatment.

Adult↗
PubMed1982

The origin of a cathode-migrating creatine kinase found in serum from a cancer patient.

The origin of an atypical creatine kinase (CK, ATP:creatine N-phosphotransferase, EC 2.7,3.2) migrating cathodic to the MM position found in the serum of a cancer patient was studied. The electrophoretic mobility of the atypical CK is similar to that of the fast-moving cathodal mitochondrial CK. The relative molecular mass was estimated to be approximately 350000, and was similar to that of the fast-moving cathodal mitochondrial CK. The atypical CK reacted with anti-human mitochondrial CK antibody. It is therefore suggested that the atypical CK is of mitochondrial origin. After incubation in 2 mol/l urea, the enzyme was converted into a new form migrating to the MM position. The conversion was observed in liver mitochondrial CK but not heart mitochondrial CK. The residual CK activity after heating at 56 degrees C for 60 s was 77%, and the apparent Km value for creatine phosphate at 30 degrees C was about 0.27 mmol for the atypical CK. These characteristics were very similar to those of the liver mitochondrial CK, because the data from the enzyme determined at the same time were 75% for residual enzyme activity to heat, and 0.24 mmol for apparent Km value. Therefore liver mitochondria are suggested to be the source of the atypical CK.

Adult↗
PubMed1982

Characteristics of mitochondrial creatine kinases from normal human heart and liver tissues.

Mitochondrial creatine kinases (CKs, ATP:creatine N-phosphotransferases, EC 2.7.3.2) were isolated from normal human heart and liver, and their characteristics were compared. The electrophoretic patterns of the extracted enzymes exhibited two forms both migrating cathodic to CK-MM. The fast-moving cathodal form is the major form and the slow-moving cathodal form is the minor one. Incubation of the heart mitochondrial CK at 37 degrees C in normal human serum for 7 h and of the liver mitochondrial CK at 26 degrees C for 1 h in 2 mol/l urea, converted the fast-moving form into the slow-moving one, and finally into a third form migrating in the MM position. The relative molecular masses were estimated to be approximately 350,000 for the major form, and 80,000 for the minor and the third forms. The electrophoretic mobility and molecular weight of the third form were identical to those of the CK-MM; however, the third form was distinguished from CK-MM by its different antigenicity. Thus, three forms were ultimately recognized as mitochondrial CKs by electrophoretic mobilities and molecular weights. The liver mitochondrial CK reacted with anti-human heart mitochondrial CK antibody, thus these two isoenzymes could not be discriminated by their antigenicities. The liver mitochondrial CK was more stable to heat and had higher apparent affinity for creatine phosphate than the heart mitochondrial CK.

Antibody Formation↗