Purification of Aldolase C from Rat Brain and Hepatoma

Hatzfeld, Antoinette; Élion, Jacques; Mennecier, F.; Schapira, F

doi:10.1111/j.1432-1033.1977.tb11638.x

Cited by 9 publications

(3 citation statements)

References 35 publications

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“…3). This result coincides with those described by others [20,221 and supports the fact that aldolase C was detected in fetal liver and hepatoma [6]. The level of the mRNA expressed in liver of the 14-day embryo was less than 2% of that expressed in adult brain.…”

Section: G C C C G T C C a T G T C T G A G G A T G G A G A A A C T A supporting

confidence: 92%

The structure of brain‐specific rat aldolase C mRNA and the evolution of aldolase isozyme genes

Kukita

Mukai

Miyata

et al. 1988

European Journal of Biochemistry

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The cDNA clones for rat aldolase C mRNA having the nearly complete length were isolated from a rat brain cDNA library and sequenced. The nucleotide sequence of pRAC2-1, a cDNA clone having the largest cDNA insert, indicates that the cDNA is composed of a 105-base-pair 5'-noncoding sequence, a 1089-base-pair codingsequence and a 382-base-pair 3'-noncoding sequence. The amino acid sequence of aldolase C deduced from a possible open reading frame was composed of 362 residues having a relative molecular mass of 39164 excluding the initiating methionine, one amino acid shorter than aldolases A and B. The length of aldolase C mRNA was 1750 residues, somewhat longer than that of the aldolase A and B transcripts. The aldolase C mRNA was distributed mainly in the brain, some in ascites hepatoma and fetal liver.Comparison of the amino acid sequences of rat aldolase C with those for rat aldolase A and B [Joh et al. (1985) Gene 39, 17 -24; Tsutsumi et al. (1984) J. Biol. Chem. 259, 14 572 -14 5751, which have been determined previously, shows the existence of highly conserved stretches of amino acid among the three isozymic forms throughout their sequences. The extent of the homology between aldolases A and C is 81%, while those between aldolases A and B, and B and C are 70%, respectively. The analysis of amino acid substitution among aldolases A, B and C from several species suggests that the isozyme genes diverged much earlier than animal species appeared and that the aldolase C gene has evolved from the aldolase A gene after aldolase A and B genes diverged.Vertebrate fructose-l,6-bisphosphate aldolase, a glycolytic enzyme, comprises three types of isozyme: aldolase A (muscle type), B (liver type), and C (brain type) [l]. These three aldolases have nearly the same molecular size, but differ in substrate specificity and electrophoretic mobility from each other. The expression of isozyme in various tissues and the change of the isozymic forms during development and carcinogenesis have been extensively studied in several species Aldolase C is mainly expressed in brain [3], some in fetal liver [4] and ascites hepatoma along with the expression of aldolase A [5, 61, and at a very low level in skeletal muscle, heart muscle and spleen [3]. Although aldolase C was suggested to be localized in neuron and glial cells of brain by immunohistochemical methods [7], it is not known, at present, why its expression is confined within brain and how the expression is regulated during development of the brain.To date, in order to elucidate the molecular mechanism of aldolase gene expression, we and other groups have determined the nucleotide sequences of cDNA and genomic genes of aldolase A and B of human [8 -121, rat [13 -171, rabbit [18] and chicken [19]. These studies have shed light on the structure and expression of aldolase isozyme A and B genes. Recently, partial sequences of aldolase C cDNA clones of mouse [20] and rat [21] and a nearly complete sequence of genomic clones of human aldolase C [22] have been reported. PI.

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Section: G C C C G T C C a T G T C T G A G G A T G G A G A A A C T A supporting

confidence: 92%

The structure of brain‐specific rat aldolase C mRNA and the evolution of aldolase isozyme genes

Kukita

Mukai

Miyata

et al. 1988

European Journal of Biochemistry

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“…Under the conditions used with the 43 kDa membrane protein, bovine brain cytosolic aldolase was phosphorylated by NDP kinase primarily on an aspartic residue. The site of phosphorylation seems to be Asp-319 on aldolase C. Aldolase C is found mostly in the brain but it is found in low levels in other tissues [28] and in some types of tumour [34][35][36].…”

Section: Discussionmentioning

confidence: 98%

Histidine to aspartate phosphotransferase activity of nm23 proteins: phosphorylation of aldolase C on Asp-319

WAGNER¹,

VU²

2000

Biochem. J.

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nm23 genes have been implicated in the suppression of tumour metastasis and cell motility; however, the biochemical mechanisms for these suppressions are not known. We have previously described the transfer of phosphate from the catalytic histidine residues of nm23 proteins to an aspartic or a glutamic residue on one or more 43 kDa proteins in detergent extracts of bovine brain membranes. To gain a better understanding of this transferase activity, we partly purified this 43 kDa protein and identified aldolases A and C as the major 43 kDa proteins present in the preparation. Aldolase was purified from brain cytosol; its phosphorylation by rat liver nm23 proteins and by recombinant human nm23-H1 was examined. The site of phosphorylation was identified as Asp-319 on aldolase C. The equivalent residue on aldolase A, a glutamic residue, was not phosphorylated. Aldolase C was rapidly phosphorylated by wild-type nm23-H1 but was not phosphorylated, or was phosphorylated very slowly, by either nm23-H1(P96S) or nm23-H1(S120G), mutants of nm23-H1 that do not suppress cell motility. This is the first identification of a protein that is phosphorylated on an aspartic residue by nm23 proteins. The sequence around Asp-319 of aldolase C has some similarities to those around the histidine residues on ATP-citrate lyase and succinic thiokinase that are phosphorylated by nm23 proteins.

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“…We shall first consider breast cancer. Aldolase B activity in cancer tissue extracts has been found to be generally quite low; and even in the tumour specimens where an appreciable amount of aldolase B is detected, it is well known that the aldolase isoenzyme pattern is shifted from B-dominant (liver or adult type) to A-dominant (muscle or foetal type) in the process of malignant transformation (Hatzfeld et al, 1978). Thus, breast cancer per se could be a factor, but may not be the critical one, for stimulation of the aldolase B-sensitized lymphocyte.…”

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confidence: 99%

Human monoclonal antibody recognizing liver-type aldolase B

Hibi¹,

Arii²,

Iizumi³

et al. 1986

Biochemical Journal

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A human hybridoma clone (4E3) has been established by fusing lymphocytes from a lymph node taken from a breast cancer patient and human lymphoblastoid cells, LICR-LON-HMy2, by the poly(ethylene glycol) method. 4E3 has been stabilized and continued to secrete IgMk antibody into culture medium (greater than 10 micrograms/ml) for over 1 year. The following characteristics of the antigen strongly suggested that 4E3 recognizes liver-type aldolase B (EC 4.1.2.13): the Mr of the native molecule is 160,000 and that of the subunit is 40,000, and thus it has a tetrameric structure of identical subunits; the antigen is abundant in the liver and kidney of human, mouse and rabbit, and is localized by immunohistochemical methods in the cytoplasm of hepatocytes and in the proximal tubules of the kidney; the antigen is precipitable by 50-80% saturation with (NH4)2SO4; the antigen shows charge-dependent heterogeneity on DEAE-cellulose chromatography. To confirm this notion, aldolase B was purified to homogeneity from the liver of human, mouse and rabbit by phosphocellulose chromatography. During the chromatographic purification, the antigen activity as assayed by enzyme-linked immunosorbent assay (e.l.i.s.a.) was superimposed on the enzymic activity of aldolase. Furthermore, monoclonal antibody 4E3 strongly reacted with purified aldolase B in SDS/polyacrylamide-gel electrophoresis followed by Western blotting and also in e.l.i.s.a. using microplates coated with purified enzyme. The reaction between aldolase B and 4E3 activated the human complement system as assessed by the attachment of C3 to the immune complex of aldolase B and 4E3.

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Purification of Aldolase C from Rat Brain and Hepatoma

Cited by 9 publications

References 35 publications

The structure of brain‐specific rat aldolase C mRNA and the evolution of aldolase isozyme genes

The structure of brain‐specific rat aldolase C mRNA and the evolution of aldolase isozyme genes

Histidine to aspartate phosphotransferase activity of nm23 proteins: phosphorylation of aldolase C on Asp-319

Human monoclonal antibody recognizing liver-type aldolase B

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