Molecular cloning of human protein 4.2: a major component of the erythrocyte membrane.

Sung, Lanping Amy; Chien, Shu; Chang, Long Sheng; Lambert, Karel J.; Bliss, Susan; Bouhassira, Eric E.; Nagel, Ronald L.; Schwartz, Robert S.; Rybicki, Anne C.

doi:10.1073/pnas.87.3.955

Cited by 61 publications

(32 citation statements)

References 38 publications

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“…The most 5' sequences of the two transcripts obtained were identical (verifying that they are derived from a single gene) and extended to nucleotide -214, numbering backwards from the translation start site. However, this position was 12 nucleotides downstream of the most 5' sequence generated by Sung et al (7), which extended to nucleotide -226. This apparent difference in the 5' origin of the band 4.2 transcript likely reflects a slight degradation of our reticulocyte RNA and in the following discussion we assume that the transcription start site is at nucleotide -226 (marked by an arrow in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…6; see also ref. 7). We found that the amino acid sequence of human erythrocyte band 4.2 has homology with two transglutaminases, guinea pig liver transglutaminase, and the a subunit of human coagulation factor XIII.…”

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confidence: 90%

“…Our genomic data show that band 4.2 exon I contains 5' noncoding sequence, the translation start site, and 99 nucleotides encoding 33 amino acids. These 33 amino acids are identical to the first 33 amino acids of the larger band 4.2 transcript (7). The 3'-most 90 nucleotides of exon I, coding for 30 amino acids, must be removed by splicing to generate the smaller transcript, coding for the 691-amino acid protein (see Fig.…”

Section: Methodsmentioning

confidence: 99%

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Organization of the gene for human erythrocyte membrane protein 4.2: structural similarities with the gene for the a subunit of factor XIII.

Korsgren

Cohen²

1991

Proc. Natl. Acad. Sci. U.S.A.

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Human erythrocyte band 4.2 is a major membrane-associated protein with an important, but still undefined, role in erythrocyte survival. We previously sequenced the complete cDNA for band 4.2 and showed that the protein has a strong sequence identity with the transglutaminase family of proteins but lacks transglutaminase activity. Here we have analyzed the genomic organization of band 4.2. The band 4.2 gene is =20 kilobases, consisting of 13 exons and 12 introns. Reticulocytes contain two different sized messages for band 4.2, and our results show that the major, smaller, message is produced by alternative splicing within band 4.2 exon I. The upstream region of the gene has several prospective promoter elements arranged in a pattern similar to that of two other erythroid genes, (3-globin and porphobilinogen deamiase.Alignment of the band 4.2 amino acid sequence with that of the a subunit of human coagulation factor XI and division of the sequences into exons reveal a remarkable correspondence, and in most cases identity, in the sizes of the paired exons. Moreover, each corresponding intron of the two genes is of an identical splice junction class. These and other similarities suggest that the gene for band 4.2 is closely related to and possibly derived from that for the a subunit of factor XIII and that the proteins may share common structural and functional properties.Human erythrocyte band 4.2 is a major membrane-associated protein of 72 kDa and is present at ==200,000 copies per cell (1, 2). Band 4.2 is clearly important for normal erythrocyte function since patients whose erythrocytes are deficient in or lack band 4.2 are anemic due to accelerated erythrocyte destruction and have abnormally shaped, possibly fragile, erythrocytes (3-5). In spite of this, however, the exact function of band 4.2 in the erythrocyte remains unclear. We have recently cloned and sequenced the full-length cDNA for band 4.2 (ref. 6; see also ref. 7). We found that the amino acid sequence of human erythrocyte band 4.2 has homology with two transglutaminases, guinea pig liver transglutaminase, and the a subunit of human coagulation factor XIII. The region of most striking identity includes a 49-amino acid domain of band 4.2, which is 69% and 51% identical to the corresponding domains of guinea pig liver transglutaminase and of the a subunit of factor XIII, respectively, and which spans the regions containing the active sites (8, 9) of both enzymes. However, band 4.2 has no demonstrable crosslinking activity when tested in vitro (6).In principle, the sequence similarities of band 4.2 with the transglutaminases could have arisen as the result of convergent evolution, in which each class of proteins evolved common, but as yet undetermined, functions independently, or, more likely, by divergent evolution, in which case the proteins would be expected to have arisen from a common ancestral gene. This question can be resolved by comparing the organization of the gene for band 4.2 with those of the transglutaminases, since a similar gen...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 90%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Organization of the gene for human erythrocyte membrane protein 4.2: structural similarities with the gene for the a subunit of factor XIII.

Korsgren

Cohen²

1991

Proc. Natl. Acad. Sci. U.S.A.

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“…On the carboxy-terminal side, aromatic residues are usually present, except for a Cys residue in human TGase 4 (Grant et al, 1994). The enzymatically inactive P4.2 isoform has an aromatic residue (Sung et al, 1990) at the amino-terminal and negatively charged residues at the carboxyterminal sides. The essential catalytic active site Cys-272 residue is located just downstream of Arg268-Tyr269, which explains the high degree of conservation in the enzymatically active isoforms.…”

Section: The Function Of Cis Peptide Bonds In Tg Asementioning

confidence: 99%

A model for the reaction mechanism of the transglutaminase 3 enzyme

Ahvazi

Steinert²

2003

Exp Mol Med

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A bstractTransglutam inase enzym es (TG ases) catalyze the calcium dependent formation of an isopeptide bond betw een protein-bound glutam ine and lysine substrates. Previously w e have show n that activated TG ase 3 acquires tw o additional calcium ions at site tw o and three. The calcium ion at site three results in the opening of a channel. A t this site, the channel opening and closing could m odulate, depending on which m etal is bound. Here we propose that the front of the channel could be used by the tw o substrates for enzym e reaction. W e propose that the glutam ine substrate is directed from Trp236 into the enzym e, shown by m olecular docking. Then a lysine substrate approaches the opened active site to engage Trp327, leading to form ation of the isopeptide bond. Further, direct com parisons of the structures of TG ase 3 w ith other TG ases have allow ed us to identify several residues that m ight potentially be involved in generic and specific recognition of the glutam ine and lysine substrates.Keywords: calcium ions; protein structure; residue specificity; TGase; TGase mechanism; X-ray crystallography IntroductionTransglutaminases (TGase; protein-glutamine: amine γ-glutamyl-transferase) are a family of calcium-dependent acyl-transfer enzymes that are widely expressed in biota (Folk and Chung, 1985;Greenberg et al., 1991;Melino et al., 1998;Melino et al., 2000;Fesus and Piacentini, 2002). Each of the eight active human TGase enzyme isoforms can activate a protein-bound Gln residue to form a thiol-acyl enzyme intermediate, which is attacked by a second nucleophilic substrate to accomplish the two-step reaction. The reaction leads to the formation of an isopeptide bond between two proteins and the covalent incorporation of polyamine into protein (Folk and Chung, 1985;Greenberg et al., 1991). The nucleophile may be: water, so that the activated Gln residue is deamidated to a Glu; a polyamine, so that a mono-substituted adduct or bi-substituted cross-link is formed; an alcohol, particularly the ω-hydroxyl group of certain long-chain ceramides expressed in mammalian epidermis, to form an ester; and perhaps most commonly, an ε-NH2 group of a protein bound Lys residue, resulting in the formation of an N ε -(γ-glutamyl)lysine isopeptide cross-link Nemes et al., 2000). This stabilizes macromolecular protein complexes. Typically, TGases recognize the generic sequence motif Gln-Gln*-Val for the first step of the reaction (where Gln* represents the targeted residue), although different isoforms display specificity as to which substrates bearing the motif may approach the enzyme . Anecdotal data suggest less specificity for Lys substrates (Tarcsa et al., 1998;Candi et al., 1999;.The three dimensional structures of four TGases have now been reported, i.e. human factor XIIIa (hfXIIIa) ), TGase 2 (Liu et al., 2002, TGase 3 enzymes (Ahvazi et al., 2002;2003), and a fish enzyme (fTG, equivalent to mammalian TGase 2, Noguchi et al., 2001). All consist of four domains that are similar in organization and fold (Figure 1...

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“…The transamidating function of the protein seems to be switched to that of receptor signaling. The membrane-skeletal protein, band 4.2, of human erythrocytes also contains sequence similarities near the active center region of TGases without, however, containing the catalytically important cysteine residue of the enzyme (51,52). No cross-reacting antibodies for the 4.2 protein and TGase have been reported.…”

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confidence: 99%