N-terminal acetylation of annexin A2 is required for S100A10 binding

Nazmi, Ali Reza; Ozorowski, Gabriel; Pejic, Milena; Whitelegge, Julian P.; Gerke, Volker; Luecke, Hartmut

doi:10.1515/hsz-2012-0179

Cited by 31 publications

(28 citation statements)

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“…Bacterial expression of AnxA2 and its mutants was achieved by using a system that faithfully executed the posttranslational N-terminal modifications seen in mammalian cells, i.e. removal of the starting methionine and acetylation of the new N-terminal serine residue (13). Correct N-terminal processing of full-length AnxA2 enabled efficient S100A10 binding and thus correct reconstitution of the heterotetrameric AnxA2-S100A10 complex (herein referred to as A2t).…”

Section: Purification and Labeling Of Recombinant Anxa2 Derivatives-mentioning

confidence: 99%

“…Thus, chemical modification of at least one of the exposed cysteines in AnxA2 (Cys-9, Cys-133, Cys-262) or S100A10 (Cys-61, Cys-82) interferes with complex formation. Therefore, to label A2t, the complex was first reconstituted from the purified individual subunits using established procedures (13) and then reacted with AlexaFluor 568 C 5 -maleimide. This yielded a fluorescently labeled complex in which both subunits were modified (Fig.…”

Section: Purification and Labeling Of Recombinant Anxa2 Derivatives-mentioning

confidence: 99%

“…The S100A10 binding site is restricted to the first 12 N-terminal residues of AnxA2, which form an amphipathic ␣-helix that is stabilized by posttranslational N-terminal acetylation (11)(12)(13). Both monomeric AnxA2 on endosomes and A2t at the plasma membrane are thought to function by organizing membrane domains in conjunction with an underlying actin cytoskeleton (14 -17).…”

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confidence: 99%

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Lipid Segregation and Membrane Budding Induced by the Peripheral Membrane Binding Protein Annexin A2*

Drücker

Pejic²,

Galla

et al. 2013

Journal of Biological Chemistry

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100

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Background: Annexin A2 is a phospholipid binding protein implicated in endo-and exocytosis. Results: Annexin A2 induces microdomain formation and inward vesicle budding in giant unilamellar vesicles. Conclusion: Specific lipid clustering and vesicle budding is triggered by annexin A2. Significance: Membrane binding by the cytosolic protein annexin A2 can induce the formation of microdomains rich in cholesterol and phosphatidylinositol 4,5-bisphosphate.

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Section: Purification and Labeling Of Recombinant Anxa2 Derivatives-mentioning

confidence: 99%

Section: Purification and Labeling Of Recombinant Anxa2 Derivatives-mentioning

confidence: 99%

mentioning

confidence: 99%

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Lipid Segregation and Membrane Budding Induced by the Peripheral Membrane Binding Protein Annexin A2*

Drücker

Pejic²,

Galla

et al. 2013

Journal of Biological Chemistry

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100

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“…It can exist in two physical states, as a monomer that is found predominantly in the cytosol and on early endosomes (5), and as a heterotetrameric complex (A2t) with its protein ligand S100A10 that is present in the subplasmalemmal region (6). S100A10, a member of the S100 family of small dimeric, EF-hand type Ca 2þ -binding proteins binds to the first 12 N-terminal residues of AnxA2, which form an amphipathic a-helix stabilized by posttranslational acetylation (7)(8)(9).…”

Section: Introductionmentioning

confidence: 99%

Cooperative Binding of Annexin A2 to Cholesterol- and Phosphatidylinositol-4,5-Bisphosphate-Containing Bilayers

Drücker¹,

Pejic²,

Grill³

et al. 2014

Biophysical Journal

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Biological membranes are organized into dynamic microdomains that serve as sites for signal transduction and membrane trafficking. The formation and expansion of these microdomains are driven by intrinsic properties of membrane lipids and integral as well as membrane-associated proteins. Annexin A2 (AnxA2) is a peripherally associated membrane protein that can support microdomain formation in a Ca(2+)-dependent manner and has been implicated in membrane transport processes. Here, we performed a quantitative analysis of the binding of AnxA2 to solid supported membranes containing the annexin binding lipids phosphatidylinositol-4,5-bisphosphate and phosphatidylserine in different compositions. We show that the binding is of high specificity and affinity with dissociation constants ranging between 22.1 and 32.2 nM. We also analyzed binding parameters of a heterotetrameric complex of AnxA2 with its S100A10 protein ligand and show that this complex has a higher affinity for the same membranes with Kd values of 12 to 16.4 nM. Interestingly, binding of the monomeric AnxA2 and the AnxA2-S100A10 complex are characterized by positive cooperativity. This cooperative binding is mediated by the conserved C-terminal annexin core domain of the protein and requires the presence of cholesterol. Together our results reveal for the first time, to our knowledge, that AnxA2 and its derivatives bind cooperatively to membranes containing cholesterol, phosphatidylserine, and/or phosphatidylinositol-4,5-bisphosphate, thus providing a mechanistic model for the lipid clustering activity of AnxA2.

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“…The N-terminus of AnxA2 binds into each of the two hydrophobic pockets of S100A10, forming a symmetric 2:2 complex of two AnxA2 molecules bound to a dimer of S100A10 (Ré ty et al, 1999). A requirement for this strong interaction is posttranslational modification of AnxA2, namely the removal of Met1 and acetylation of Ser2 at the new N-terminus (Nazmi et al, 2012;Becker et al, 1990). (p11) 2 (AnxA2) 2 is often localized near the plasma membrane, and this symmetric platform is proposed to be involved in the bridging of membrane vesicles and trafficking of proteins to the plasma membrane (van de Graaf et al, 2003;Umbrecht-Jenck et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Structure of a C-terminal AHNAK peptide in a 1:2:2 complex with S100A10 and an acetylated N-terminal peptide of annexin A2

Ozorowski

Milton

Luecke

2012

Acta Crystallogr D Biol Crystallogr

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AHNAK, a large 629 kDa protein, has been implicated in membrane repair, and the annexin A2–S100A10 heterotetramer [(p11)2(AnxA2)2)] has high affinity for several regions of its 1002‐amino‐acid C‐terminal domain. (p11)2(AnxA2)2 is often localized near the plasma membrane, and this C2‐symmetric platform is proposed to be involved in the bridging of membrane vesicles and trafficking of proteins to the plasma membrane. All three proteins co‐localize at the intracellular face of the plasma membrane in a Ca2+‐dependent manner. The binding of AHNAK to (p11)2(AnxA2)2 has been studied previously, and a minimal binding motif has been mapped to a 20‐amino‐acid peptide corresponding to residues 5654–5673 of the AHNAK C‐terminal domain. Here, the 2.5 Å resolution crystal structure of this 20‐amino‐acid peptide of AHNAK bound to the AnxA2–S100A10 heterotetramer (1:2:2 symmetry) is presented, which confirms the asymmetric arrangement first described by Rezvanpour and coworkers and explains why the binding motif has high affinity for (p11)2(AnxA2)2. Binding of AHNAK to the surface of (p11)2(AnxA2)2 is governed by several hydrophobic interactions between side chains of AHNAK and pockets on S100A10. The pockets are large enough to accommodate a variety of hydrophobic side chains, allowing the consensus sequence to be more general. Additionally, the various hydrogen bonds formed between the AHNAK peptide and (p11)2(AnxA2)2 most often involve backbone atoms of AHNAK; as a result, the side chains, particularly those that point away from S100A10/AnxA2 towards the solvent, are largely interchangeable. While the structure‐based consensus sequence allows interactions with various stretches of the AHNAK C‐terminal domain, comparison with other S100 structures reveals that the sequence has been optimized for binding to S100A10. This model adds new insight to the understanding of the specific interactions that occur in this membrane‐repair scaffold.

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N-terminal acetylation of annexin A2 is required for S100A10 binding

Cited by 31 publications

References 0 publications

Lipid Segregation and Membrane Budding Induced by the Peripheral Membrane Binding Protein Annexin A2*

Lipid Segregation and Membrane Budding Induced by the Peripheral Membrane Binding Protein Annexin A2*

Cooperative Binding of Annexin A2 to Cholesterol- and Phosphatidylinositol-4,5-Bisphosphate-Containing Bilayers

Structure of a C-terminal AHNAK peptide in a 1:2:2 complex with S100A10 and an acetylated N-terminal peptide of annexin A2

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