The calcium binding sites in human annexin V by crystal structure analysis at 2.0 A resolution Implications for membrane binding and calcium channel activity

Huber, Robert; Schneider, Monika; Mayr, Irmgard; Römisch, Jürgen; Pâques, Eric-P.

doi:10.1016/0014-5793(90)81428-q

Cited by 235 publications

(175 citation statements)

References 30 publications

Supporting

Mentioning

164

Contrasting

Unclassified

Order By: Relevance

“…Similar Ca a~ channel activities were recently reported for annexin V [32]. Data from the analysis of the crystal ~ ,,"),, of human ann~,vin V support the hypothesis that this protein might have Ca"* channel activities [33].…”

Section: 5supporting

confidence: 83%

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

et al. 1992

View full text Add to dashboard Cite

The distribution of annexin V isoforms (CaBP33 and CaBP37) and of annexin VI in bovine lung, heart, and brain subfractions was investigated with special reference lo the fractions of these proteins which are membrane.bound. In addition to EGTA.extmetable pools of the above proteins, membranes from lung, heart, and brain contain EGTA-resistant annexins V and VI which can be solubilizcd whh detergents (Triton X-100 or Triton X-114). A strong base like Na:CO~, which is usually effective in extracting peripheral membrane proteins, only partially solubilizes the membranebound. EGTA-resistant annexins analyzed here. Also, only 50-60% of the Triton X-114-soluble ann¢xins partition in the aqueous phase, the remaining fraetion~ being recovered in the detergent-rich phase. Altogether, these findings suggest that, by an as ),et unknown mechanism, following Ca:'-dependent association of annexin V isoforms and annexin VI with membranes, substantial fractions of tbe~e proteins remain bound to membranes in a Ca:*-indepcndent way and behave like integral membrane proteins. These results further support the possibility that the above annexins might play a role in membrane trafficking and/or in the regulation of the structural organization of membranes.Annexin V (CaBP33 and CaBP37); Annexin Vl; Membrane; Binding; Calcium; Lun~g; Heart; Brain I, INTRODUCTIONProteins of the annexin family share the ability to bind to acidic phospholipids and to natural membranes in the presence of Ca:* (for reviews see [1][2][3]). Each annexin is made of an N-terminal tail of variable length and unique to individual annexins, which is supposed to play a role in the diversification of the biological functions of single species, and of a core. This latter is made of four, in the ease of the 32-37 kDa annexins, or eight, in the case of the 67-73 kDa annexins, internal repeats 70 residues in length, each of which contains a highly conserved consensus sequence, the endonexin fold, which is suggested to take part in the coordination of binding of both Ca 2+ and phospholipids [3][4][5]. Unlike the Ca-'+-binding pro~:eins of the EF-hand type, Ca-'*-binding to annexins does not induce the exposure of hydrophobic domains; rather~ Ca 2. would cross-bridge any annexin to the negatively charged headgroups of acidic phospholipids and/or certain annexins to target proteins [1][2][3]. Given the above model, it is expected that chelation of Ca 2÷ will result in the complete reversal of annexin binding. While this has been proven true whenever annexin binding to liposomes or proteins had been investigated in reconstitution experiments in vitro [6][7][8][9][10][11][12][13][14][15], different results were obtained with certain annexins whenever tissue or cell subfractionation was used to investigate the binding of endogenous annexins to natural membranes. So, fractions ofannexins I, 1I, IV-VII were reported to exist in several cell types in a membrane-bound form, to resist extraction with EGTA, and to require detergents for their solubilization [10,[16][17][18][19][2...

show abstract

Section: 5supporting

confidence: 83%

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

et al. 1992

View full text Add to dashboard Cite

show abstract

“…Here we have generated site-specifically mutated annexin II molecules which allow the precise description of the H28 epitope. Our results show that in the region of the H28 epitope, the three-dimensional folding of annexin II must be highly similar to that of annexin V, which was recently determined by X-ray diffraction [8,12] …”

Section: Phosphorylationsupporting

confidence: 62%

“…Thus it seems that a conformational change induced by phosphorylation is responsible for the altered Ca'+/lipid-binding properties. Whether this conformational change involves the pore, which transverses the annexin molecule and is thought to be a voltage dependent Ca* ' channel [8,12], is not known. It seems however important that all regulatory elements of annexin II, the phosphorylation sites for kinase C and pp60"" at Ser23 and Tyr2', as well as the pl 1 binding site at residues 1 to 14 are probably situated in close proximity to the entrance of the pore.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a discontinuous epitope on annexin II by site‐directed mutagenesis

1991

View full text Add to dashboard Cite

Recombinant annexin II mutants were generated to identify amino acids involved in the formation of the discontinuous cpitope of the monoclonal antibody H28. Analysis of the various mutant proteins by immunoblotting and enzyme-linked immunosorbent assay revealed that residues Lysz7, Arg2, GW, and Arg7 are indispensable for H28 reactivity. Residues in equivalent positions are also in close proximity in the recently determined X-ray structure of annexin V, a different member of the same family of Ca'+/lipid-binding proteins. Thus annexins II and V show a similar threedimensional folding in this region of the molecule. Consequently, !he Ca** binding sites and the residues phosphorylated by pp601~ (Tyti') and protein kinase C (Setis) most likely reside on opposite sides of the annexin II molecule.

show abstract

“…In the individual repeat the endonexin fold covers helix b and the preceding interhelical loop between helices a and b which contains the highly conserved glycine and threonine residues (positions 4 and 5 of the endonexin fold). Three Ca2+-binding sites in the annexin V crystals, which were identified in repeats 1, 2, and 4, are of particular interest since they differ from well characterized Ca" sites such as the helix-loop-helix motif of the EF-hand (Huber et al, 1990b). The 'annexin-type' Ca2+-binding sites are formed in part by the loop of the individual endonexin folds.…”

mentioning

confidence: 99%

“…Additional coordination of the Ca2+ ion is provided by the carboxyl oxygens of a glutamate or aspartate residue located in helix d of the repeat (40 residues C-terminal from the conserved glycine of the endonexin fold). In phospholipidbound annexins, another Ca2+-coordination site is thought to be provided by the phosphoryl moiety of the phospholipid (Huber et al, 1990b). This takes into account the mutual influence of Ca2+ and phospholipid binding by annexins; i.e.…”

mentioning

confidence: 99%

Mapping of three unique Ca²⁺‐binding sites in human annexin II

Jost

Thiel

Weber

et al. 1992

European Journal of Biochemistry

View full text Add to dashboard Cite

Site-directed mutagenesis was employed to map and characterize Ca2 + -binding sites in annexin 11, a member of the annexin family of Ca2+-and phospholipid-binding proteins which serves as a major cellular substrate for the tyrosine kinase encoded by the src oncogene. Several single amino acid substitutions were introduced in the human annexin I1 and the various mutant proteins were scored for their affinity towards Ca2+ in different assays. The data support our previous finding [Thiel, C., Weber, K. and Gerke V. (1991) J . Biol. Chem. 266, 14732-147391 that a Ca2+-binding site is present in the third of the four repeat segments whch comprise the 33-kDa protein core of annexin 11. In addition to Gly206 and Thr207, which are localized in the highly conserved endonexin fold of the third repeat, Glu246 is involved in the formation of this site. Thus the architecture of this Ca2+-binding site in solution is very similar, if not identical, to that of Ca2+ sites identified recently in annexin V crystals [Huber, R., Schneider, M., Mayr, I., Romisch, J. and Paques, E.-P. (1990) FEBS Lett. 275,[15][16][17][18][19][20][21]. In addition to the site in repeat 3, we have mapped sites of presumably similar architecture in repeats 2 and 4 of annexin 11. Again, an acidic amino acid which is located 40 residues C-terminal to the conserved glycine at position 4 of the endonexin fold is indispensable for highaffinity Ca2+ binding: Asp161 in the second and Asp321 in the fourth repeat. In contrast, repeat 1 does not contain an acidic amino acid at a corresponding position and also shows deviations from the other repeats in the sequence surrounding the conserved glycine. These results on annexin I1 together with the crystallographic information on annexin V reveal that annexins can differ in the position of the Ca2+ sites. Ca2+-binding sites of similar structure are present in repeats 2, 3, and 4 of annexin I1 while in annexin V they occur in repeats 1, 2, and 4. We also synthesized an annexin I1 derivative with mutations in all three Ca2+ sites. This molecule shows a greatly reduced affinity for the divalent cation. However, it is still able to bind Ca2+, indicating the presence of (an) additional Ca2 + site(s) of presumably different architecture.The annexins form a multigene family of Ca2+-dependent membrane-binding and phospholipid-binding proteins. Although the precise biological function of the annexins is not known, their biochemical properties imply that the proteins are involved in membrane phenomena, e.g. membrane fusion events during exocytosis, membrane-cytoskeleton linkage, membrane channel formation (for review see Klee, 1988;Moss et al., 1991). In addition to common biochemical properties the annexins share a characteristic structural element, the annexin repeat. This segment of 70-80 amino acids is repeated four (32 -39-kDa annexins) or eight times (68-kDa annexin) along the polypeptide chain of the individual annexin. The annexin repeats show intramolecular as well as intermolecular sequence similarities which are par...

show abstract

The calcium binding sites in human annexin V by crystal structure analysis at 2.0 A resolution Implications for membrane binding and calcium channel activity

Cited by 235 publications

References 30 publications

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

Characterization of a discontinuous epitope on annexin II by site‐directed mutagenesis

Mapping of three unique Ca²⁺‐binding sites in human annexin II

Contact Info

Product

Resources

About

The calcium binding sites in human annexin V by crystal structure analysis at 2.0 A resolution Implications for membrane binding and calcium channel activity

Cited by 235 publications

References 30 publications

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

Membrane‐bound annexin V isoforms (CaBP33 and CaBP37) and annexin VI in bovine tissues behave like integral membrane proteins

Characterization of a discontinuous epitope on annexin II by site‐directed mutagenesis

Mapping of three unique Ca2+‐binding sites in human annexin II

Contact Info

Product

Resources

About

Mapping of three unique Ca²⁺‐binding sites in human annexin II