A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin

David A. Gell, Liang Feng, Suiping Zhou, Philip D. Jeffrey, Katerina Bendak, Andrew Gow, Mitchell J. Weiss, Yigong Shi, Joel P. Mackay

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops.

Original languageEnglish
Pages (from-to)29462-29469
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number43
DOIs
Publication statusPublished - 23 Oct 2009
Externally publishedYes

Fingerprint

Proline
Hemoglobins
Proteins
Conformations
Heme
Mutagenesis
Protein Conformation
Crystallography
Mutation
Accessories
Histidine
Nuclear magnetic resonance spectroscopy
Reactive Oxygen Species
Magnetic Resonance Spectroscopy
X-Rays
Tissue
X rays

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Gell, D. A., Feng, L., Zhou, S., Jeffrey, P. D., Bendak, K., Gow, A., ... Mackay, J. P. (2009). A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin. Journal of Biological Chemistry, 284(43), 29462-29469. https://doi.org/10.1074/jbc.M109.027045

A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin. / Gell, David A.; Feng, Liang; Zhou, Suiping; Jeffrey, Philip D.; Bendak, Katerina; Gow, Andrew; Weiss, Mitchell J.; Shi, Yigong; Mackay, Joel P.

In: Journal of Biological Chemistry, Vol. 284, No. 43, 23.10.2009, p. 29462-29469.

Research output: Contribution to journalArticle

Gell, DA, Feng, L, Zhou, S, Jeffrey, PD, Bendak, K, Gow, A, Weiss, MJ, Shi, Y & Mackay, JP 2009, 'A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin', Journal of Biological Chemistry, vol. 284, no. 43, pp. 29462-29469. https://doi.org/10.1074/jbc.M109.027045
Gell, David A. ; Feng, Liang ; Zhou, Suiping ; Jeffrey, Philip D. ; Bendak, Katerina ; Gow, Andrew ; Weiss, Mitchell J. ; Shi, Yigong ; Mackay, Joel P. / A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 43. pp. 29462-29469.
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