Muscle creatine kinase isoenzyme expression in adult human brain.
Previous studies have suggested that MM creatine kinase is a muscle-specific protein and is not present in adult brain tissue. We have isolated a protein from human brain with an apparent molecular weight of 43,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis which is identical to the muscle M creatine kinase isoenzyme subunit at all 30 sequenced amino acid residues and possesses creatine kinase enzymatic activity following nondenaturing agarose-gel electrophoresis. Immunohistochemistry localizes M creatine kinase to discrete areas of adult human brain. Northern blot analysis of both total and poly(A)-selected RNA isolated from brain did not detect M creatine kinase mRNA. However, polymerase chain reaction amplification of cDNA synthesized from human placenta, heart, and brain mRNA detected M creatine kinase message in both heart and brain but not placenta which contains no detectable M creatine kinase protein. N1E115 and NS20Y, mouse neuroblastoma cell lines which have been used as models of neural cell differentiation, were found also to express MM creatine kinase. Moreover, a transiently transfected reporter gene with 4,800 base pairs of M creatine kinase upstream region fused to chloramphenicol acetyltransferase was expressed during differentiation of these neural cell lines. In summary, MM creatine kinase is present in human brain and we suggest the M creatine kinase upstream region is sufficient to modulate M creatine kinase expression in certain neuronal cells and may be regulated independently from other muscle genes.
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.
R J
RJ
Hamburg
D L
DL
Friedman
E N
EN
Olson
T S
TS
Ma
M D
MD
Cortez
C
C
Goodman
P R
PR
Puleo
M B
MB
Perryman
D000328:Adult; D000595:Amino Acid Sequence; D001921:Brain / Q000201:enzymology*; D002460:Cell Line; D003402:Creatine Kinase / Q000235:genetics* / Q000302:isolation & purification; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005784:Gene Amplification; D015870:Gene Expression; D006624:Hippocampus / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D015345:Oligonucleotide Probes; D010920:Placenta / Q000201:enzymology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012194:RNA-Directed DNA Polymerase; D013702:Temporal Lobe / Q000201:enzymology; D014162:Transfection
D007527:Isoenzymes; D015345:Oligonucleotide Probes; D004247:DNA; D003402:Creatine Kinase; D012194:RNA-Directed DNA Polymerase