scholarly journals Band 3 Tuscaloosa: Pro327----Arg327 substitution in the cytoplasmic domain of erythrocyte band 3 protein associated with spherocytic hemolytic anemia and partial deficiency of protein 4.2

Blood ◽  
1992 ◽  
Vol 80 (2) ◽  
pp. 523-529 ◽  
Author(s):  
P Jarolim ◽  
J Palek ◽  
HL Rubin ◽  
JT Prchal ◽  
C Korsgren ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Binding Capacity ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Molecular Defect ◽  
Band 3 Protein ◽  
Additional Line ◽  
Peripheral Proteins ◽  
Protein 4.2 ◽  
Partial Deficiency

Protein 4.2 is a major red blood cell (RBC) protein that interacts with the band 3 protein and with ankyrin. Inherited deficiencies of this protein are associated with spherocytic hemolytic anemia, but the molecular basis of this defect is unknown. We have studied the underlying defect in a patient with spherocytic hemolytic anemia whose RBCs had a partial (29% +/- 5%) deficiency of protein 4.2. We have first studied the binding of normal ankyrin and protein 4.2 to patient inside-out vesicles (IOVs) stripped of peripheral proteins. While the binding of ankyrin was normal, the predicted maximal binding capacity of patient IOVs for band 4.2 was 20% to 33% lower than that of control IOVs, suggesting a defect in the cytoplasmic domain of band 3 (cdb3). An additional line of evidence pointing to a possible abnormality of band 3 was an abnormal proteolytic digest of cdb3. To elucidate the underlying molecular defect, we have cloned and sequenced the cDNA coding for cdb3 from the patient. One band 3 allele was found to be normal, while clones corresponding to the other allele contained two mutations: substitution A----G in nucleotide 166, changing codon 56 from AAG to GAG (Lys----Glu), and substitution C----G in nucleotide 980, changing codon 327 from CCC to CGC (Pro----Arg). Since the Lys56--- -Glu56 substitution is found in a common asymptomatic variant of the band 3 protein designated band 3 Memphis, we conclude that either the Pro327----Arg327 substitution itself, or in combination with the band 3 Memphis polymorphism, underlies the abnormal binding of protein 4.2 to cdb3 and results in the spherocytic.

scholarly journals Band 3 Tuscaloosa: Pro327----Arg327 substitution in the cytoplasmic domain of erythrocyte band 3 protein associated with spherocytic hemolytic anemia and partial deficiency of protein 4.2

Blood ◽  
1992 ◽  
Vol 80 (2) ◽  
pp. 523-529 ◽  
Author(s):  
P Jarolim ◽  
J Palek ◽  
HL Rubin ◽  
JT Prchal ◽  
C Korsgren ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Binding Capacity ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Molecular Defect ◽  
Band 3 Protein ◽  
Additional Line ◽  
Peripheral Proteins ◽  
Protein 4.2 ◽  
Partial Deficiency

Abstract Protein 4.2 is a major red blood cell (RBC) protein that interacts with the band 3 protein and with ankyrin. Inherited deficiencies of this protein are associated with spherocytic hemolytic anemia, but the molecular basis of this defect is unknown. We have studied the underlying defect in a patient with spherocytic hemolytic anemia whose RBCs had a partial (29% +/- 5%) deficiency of protein 4.2. We have first studied the binding of normal ankyrin and protein 4.2 to patient inside-out vesicles (IOVs) stripped of peripheral proteins. While the binding of ankyrin was normal, the predicted maximal binding capacity of patient IOVs for band 4.2 was 20% to 33% lower than that of control IOVs, suggesting a defect in the cytoplasmic domain of band 3 (cdb3). An additional line of evidence pointing to a possible abnormality of band 3 was an abnormal proteolytic digest of cdb3. To elucidate the underlying molecular defect, we have cloned and sequenced the cDNA coding for cdb3 from the patient. One band 3 allele was found to be normal, while clones corresponding to the other allele contained two mutations: substitution A----G in nucleotide 166, changing codon 56 from AAG to GAG (Lys----Glu), and substitution C----G in nucleotide 980, changing codon 327 from CCC to CGC (Pro----Arg). Since the Lys56--- -Glu56 substitution is found in a common asymptomatic variant of the band 3 protein designated band 3 Memphis, we conclude that either the Pro327----Arg327 substitution itself, or in combination with the band 3 Memphis polymorphism, underlies the abnormal binding of protein 4.2 to cdb3 and results in the spherocytic.

scholarly journals Human erythrocyte protein 4.2 deficiency associated with hemolytic anemia and a homozygous 40glutamic acid-->lysine substitution in the cytoplasmic domain of band 3 (band 3Montefiore)

Blood ◽  
1993 ◽  
Vol 81 (8) ◽  
pp. 2155-2165 ◽  
Author(s):  
AC Rybicki ◽  
JJ Qiu ◽  
S Musto ◽  
NL Rosen ◽  
RL Nagel ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Hemolytic Anemia ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Structural Abnormality ◽  
Cell Deformability ◽  
Whole Cell ◽  
Rbc Membrane ◽  
Protein 4.2 ◽  
Lysine Substitution

Abstract Red blood cell (RBC) protein 4.2 deficiency is often associated with a moderate nonimmune hemolytic anemia, splenomegaly, and osmotically fragile RBCs resembling, but not identical to, hereditary spherocytosis (HS). In the Japanese type of protein 4.2 deficiency (protein 4.2Nippon), the anemia is associated with a point mutation in the protein 4.2 cDNA. In this report, we describe a patient with moderate and apparently episodic nonimmune hemolytic anemia with splenomegaly, spherocytosis, osmotically fragile RBCs, reduced whole cell deformability, and abnormally dense cells. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis analysis of the proposita's RBC membrane proteins showed an 88% deficiency of protein 4.2 and a 30% deficiency of glyceraldehyde-3-phosphate dehydrogenase (band 6). Structural and molecular analyses of the proposita's protein 4.2 were normal. In contrast, limited tryptic digestion of the proposita's band 3 showed a homozygous abnormality in the cytoplasmic domain. Analysis of the pedigree disclosed six members who were heterozygotes for the band 3 structural abnormality and one member who was a normal homozygote. Direct sequence analysis of the abnormal band 3 tryptic peptide suggested that the structural abnormality resided at or near residue 40. Sequence analysis of the proposita's band 3 cDNA showed a 232G-->A mutation resulting in a 40glutamic acid-->lysine substitution (band 3Montefiore). Allele-specific oligonucleotide hybridization was used to probe for the mutation in the pedigree, showing that the proposita was homozygous, and the pedigree members who were heterozygous for the band 3 structural abnormality were also heterozygous for the band 3Montefiore mutation. The band 3Montefiore mutation was absent in 26 chromosomes from race-matched controls and in one pedigree member who did not express the band 3 structural abnormality. In coincidence with splenectomy, the proposita's anemia was largely corrected along with the disappearance of most spherocytes and considerable improvements of RBC osmotic fragility, whole cell deformability, and cell density. We conclude that this hereditary hemolytic anemia is associated with the homozygous state for band 3Montefiore (40glutamic acid-->lysine) and a decreased RBC membrane content of protein 4.2. We speculate that band 3 structural abnormalities can result in defective interactions with protein 4.2 and band 6, and in particular, that the region of band 3 containing 40glutamic acid is involved directly or indirectly in interactions with these proteins.

scholarly journals Human erythrocyte protein 4.2 deficiency associated with hemolytic anemia and a homozygous 40glutamic acid-->lysine substitution in the cytoplasmic domain of band 3 (band 3Montefiore)

Blood ◽  
1993 ◽  
Vol 81 (8) ◽  
pp. 2155-2165 ◽  
Author(s):  
AC Rybicki ◽  
JJ Qiu ◽  
S Musto ◽  
NL Rosen ◽  
RL Nagel ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Hemolytic Anemia ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Structural Abnormality ◽  
Cell Deformability ◽  
Whole Cell ◽  
Rbc Membrane ◽  
Protein 4.2 ◽  
Lysine Substitution

Red blood cell (RBC) protein 4.2 deficiency is often associated with a moderate nonimmune hemolytic anemia, splenomegaly, and osmotically fragile RBCs resembling, but not identical to, hereditary spherocytosis (HS). In the Japanese type of protein 4.2 deficiency (protein 4.2Nippon), the anemia is associated with a point mutation in the protein 4.2 cDNA. In this report, we describe a patient with moderate and apparently episodic nonimmune hemolytic anemia with splenomegaly, spherocytosis, osmotically fragile RBCs, reduced whole cell deformability, and abnormally dense cells. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis analysis of the proposita's RBC membrane proteins showed an 88% deficiency of protein 4.2 and a 30% deficiency of glyceraldehyde-3-phosphate dehydrogenase (band 6). Structural and molecular analyses of the proposita's protein 4.2 were normal. In contrast, limited tryptic digestion of the proposita's band 3 showed a homozygous abnormality in the cytoplasmic domain. Analysis of the pedigree disclosed six members who were heterozygotes for the band 3 structural abnormality and one member who was a normal homozygote. Direct sequence analysis of the abnormal band 3 tryptic peptide suggested that the structural abnormality resided at or near residue 40. Sequence analysis of the proposita's band 3 cDNA showed a 232G-->A mutation resulting in a 40glutamic acid-->lysine substitution (band 3Montefiore). Allele-specific oligonucleotide hybridization was used to probe for the mutation in the pedigree, showing that the proposita was homozygous, and the pedigree members who were heterozygous for the band 3 structural abnormality were also heterozygous for the band 3Montefiore mutation. The band 3Montefiore mutation was absent in 26 chromosomes from race-matched controls and in one pedigree member who did not express the band 3 structural abnormality. In coincidence with splenectomy, the proposita's anemia was largely corrected along with the disappearance of most spherocytes and considerable improvements of RBC osmotic fragility, whole cell deformability, and cell density. We conclude that this hereditary hemolytic anemia is associated with the homozygous state for band 3Montefiore (40glutamic acid-->lysine) and a decreased RBC membrane content of protein 4.2. We speculate that band 3 structural abnormalities can result in defective interactions with protein 4.2 and band 6, and in particular, that the region of band 3 containing 40glutamic acid is involved directly or indirectly in interactions with these proteins.

Molecular Defect of the Band 3 Protein in Southeast Asian Ovalocytosis

1990 ◽  
Vol 323 (22) ◽  
pp. 1530-1538 ◽  
Author(s):  
Shih-Chun Liu ◽  
Sen Zhai ◽  
Jiri Palek ◽  
David E. Golan ◽  
Dominick Amato ◽  
...  
Keyword(s):  
Band 3 ◽  
Southeast Asian ◽  
Molecular Defect ◽  
Band 3 Protein

Total absence of protein 4.2 and partial deficiency of band 3 in hereditary spherocytosis

1997 ◽  
Vol 99 (3) ◽  
pp. 522-530 ◽  
Author(s):  
Akio Kanzaki ◽  
Sandrine Hayette ◽  
Laurette Morlé ◽  
Fumihide Inoue ◽  
Reiko Matsuyama ◽  
...  
Keyword(s):  
Hereditary Spherocytosis ◽  
Band 3 ◽  
Total Absence ◽  
Protein 4.2 ◽  
Partial Deficiency

scholarly journals Band 3 protein degradation by calpain is enhanced in erythrocytes of old people

1991 ◽  
Vol 275 (1) ◽  
pp. 47-52 ◽  
Author(s):  
N Schwarz-Ben Meir ◽  
T Glaser ◽  
N S Kosower
Keyword(s):  
Protein Degradation ◽  
Membrane Vesicles ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Erythrocyte Ghosts ◽  
Band 3 Protein ◽  
Enhanced Sensitivity ◽  
Age Related ◽  
The Difference ◽  
Membrane Spanning

Band 3 protein is a major erythrocyte transmembrane glycoprotein. We compared the degradation of band 3 protein by calpain I (a cytoplasmic, micromolar-Ca2(+)-requiring thiol proteinase) in the cells from old individuals (greater than 70 years old) to that in the cells from young ones (20-30 years old). In the young, little degradation of band 3 protein occurred in calpain-treated erythrocyte ghosts. In the old, significant band 3 protein degradation was found in erythrocyte ghosts treated similarly. The difference between young and old in the susceptibility of band 3 protein to calpain was retained in membrane vesicles (membranes stripped of peripheral proteins by NaOH) and in chymotrypsin-generated 60 kDa fragment (CH-60). The isolated N-terminal cytoplasmic 43 kDa fragment was degraded by calpain to a similar extent in old and in young. The separated 17 kDa membrane domain of the CH-60 and the trypsin-generated C-terminal 55 kDa membrane-spanning fragment were not degraded by calpain I in the young, nor in the old. Thus the N-terminal cytoplasmic domain is the domain degraded by calpain I. Enhanced sensitivity in the old is observed in intact band 3 protein and in CH-60, the isolated cytoplasmic domain being equally susceptible in young and old. The observed age-related enhanced sensitivity to calpain is consistent with the presence of modifications in band 3 protein and alterations in the association with the calpain-calpastatin system. Band 3 protein has several important functions, with modifications in the protein having implications for altered cell behaviour in the old individual.

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