Proteolytic cleavage sites of band 3 protein in alkali-treated membranes: Fidelity of hydropathy prediction for band 3 protein

Naotaka Hamasaki, Kenshi Okubo, Hiroyuki Kuma, Dongchon Kang, Yoshiaki Yae

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

To assess the fidelity of hydropathy prediction for band 3 protein, we determined the cleavage sites of the protein and the portions of the protein tightly bound to the membrane lipid bilayer by means of in situ proteolytic digestion. For the removal of all anticipated hydrophilic connector loops from membranes, we had to denature the band 3 protein molecule in situ by alkali treatment. When the alkali-treated membranes were digested with trypsin, chymotrypsin, and pepsin, the majority of the anticipated transmembrane portions remained in the membrane fraction. However, five anticipated transmembrane portions were released into the supernatant fraction. Thus, the first, second, third, sixth and tenth anticipated transmembrane portions, in accordance with the hydropathy prediction, were released into the supernatant with the proteolytic digestion method. This indicates that these anticipated transmembrane portions are not bound with the boundary lipids although the hydrophobicity of these portions is comparable to that of the portions experimentally remaining in the membrane fraction. It is conceivable that the membrane peptide portions of band 3 protein could be classified into at least two categories, i.e. one bound to the boundary lipids and the other free from the boundary lipids. Approximately 90% of the transmembrane domain of the band 3 protein are recovered in either the supernatant fraction or the membrane fraction. The fidelity of hydropathy prediction for polytopic membrane proteins and the nature of the membrane embedded peptide portions are discussed.

Original languageEnglish
Pages (from-to)577-585
Number of pages9
JournalJournal of Biochemistry
Volume122
Issue number3
DOIs
Publication statusPublished - Jan 1 1997

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Erythrocyte Anion Exchange Protein 1
Alkalies
Membranes
Lipids
Digestion
Peptides
Lipid bilayers
Pepsin A
Lipid Bilayers
Hydrophobicity
Membrane Lipids
Hydrophobic and Hydrophilic Interactions
Membrane Proteins
Proteins
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Proteolytic cleavage sites of band 3 protein in alkali-treated membranes : Fidelity of hydropathy prediction for band 3 protein. / Hamasaki, Naotaka; Okubo, Kenshi; Kuma, Hiroyuki; Kang, Dongchon; Yae, Yoshiaki.

In: Journal of Biochemistry, Vol. 122, No. 3, 01.01.1997, p. 577-585.

Research output: Contribution to journalArticle

Hamasaki, Naotaka ; Okubo, Kenshi ; Kuma, Hiroyuki ; Kang, Dongchon ; Yae, Yoshiaki. / Proteolytic cleavage sites of band 3 protein in alkali-treated membranes : Fidelity of hydropathy prediction for band 3 protein. In: Journal of Biochemistry. 1997 ; Vol. 122, No. 3. pp. 577-585.
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