Molecular cloning of human protein 4.2: a major component of the erythrocyte membrane.
Protein 4.2 (P4.2) comprises approximately 5% of the protein mass of human erythrocyte (RBC) membranes. Anemia occurs in patients with RBCs deficient in P4.2, suggesting a role for this protein in maintaining RBC stability and integrity. We now report the molecular cloning and characterization of human RBC P4.2 cDNAs. By immunoscreening a human reticulocyte cDNA library and by using the polymerase chain reaction, two cDNA sequences of 2.4 and 2.5 kilobases (kb) were obtained. These cDNAs differ only by a 90-base-pair insert in the longer isoform located three codons downstream from the putative initiation site. The 2.4- and 2.5-kb cDNAs predict proteins of approximately 77 and approximately 80 kDa, respectively, and the authenticity was confirmed by sequence identity with 46 amino acids of three cyanogen bromide-cleaved peptides of P4.2. Northern blot analysis detected a major 2.4-kb RNA species in reticulocytes. Isolation of two P4.2 cDNAs implies existence of specific regulation of P4.2 expression in human RBCs. Human RBC P4.2 has significant homology with human factor XIII subunit a and guinea pig liver transglutaminase. Sequence alignment of P4.2 with these two transglutaminases, however, revealed that P4.2 lacks the critical cysteine residue required for the enzymatic crosslinking of substrates.
Department of Applied Mechanics, University of California at San Diego, La Jolla 92093.
L A
LA
Sung
S
S
Chien
L S
LS
Chang
K
K
Lambert
S A
SA
Bliss
E E
EE
Bouhassira
R L
RL
Nagel
R S
RS
Schwartz
A C
AC
Rybicki
D000595:Amino Acid Sequence; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D004247:DNA / Q000235:genetics; D004910:Erythrocyte Membrane / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012313:RNA / Q000097:blood / Q000235:genetics; D015183:Restriction Mapping; D012156:Reticulocytes / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D015345:Oligonucleotide Probes; C044791:erythrocyte membrane band 4.2 protein; D012313:RNA; D004247:DNA