Trimers, Dimers of Trimers, and Trimers of Trimers Are Common Building Blocks of Annexin A5 Two-Dimensional Crystals

Oling, Frank; Bergsma-Schutter, Wilma; Brisson, Alain

doi:10.1006/jsbi.2000.4337

Cited by 111 publications

(106 citation statements)

References 26 publications

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“…stabilizing trimer subunit interactions). The crystal structure of annexin IV (Thr 6 3 Asp mutant; PDB code 1I4A) crystallized from ammonium sulfate presents the four conserved basic residues in a subtly different spatial arrangement than in annexin V. However, the cluster appears to be functionally similar in the two proteins. In annexin IV (Fig.…”

Section: Promotion Of Subunit Interactions By Basic Clustermentioning

confidence: 99%

“…The annexin-phospholipid association is of high affinity but is reversible with the removal of Ca 2ϩ . When bound to the membrane surface, annexin V molecules assemble laterally into well organized trimers and higher order arrays of trimers (6). This lateral assembly promotes spontaneous two-dimensional crystallization of annexin V on membrane surfaces, a process that is inhibited by extreme membrane surface curvature (7,8).…”

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confidence: 99%

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Interfacial Basic Cluster in Annexin V Couples Phospholipid Binding and Trimer Formation on Membrane Surfaces

Mo¹,

Campos²,

Mealy³

et al. 2003

Journal of Biological Chemistry

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Annexin V is an abundant eukaryotic protein that binds phospholipid membranes in a Ca2؉ -dependent manner. In the present studies, site-directed mutagenesis was combined with x-ray crystallography and solution liposome binding assays to probe the functional role of a cluster of interfacial basic residues in annexin V. Four mutants were investigated: R23E, K27E, R61E, and R149E. All four mutants exhibited a significant reduction in adsorption to phospholipid membranes relative to the wild-type protein, and the R23E mutation was the most deleterious. Crystal structures of wild-type and mutant proteins were similar except for local changes in salt bridges involving basic cluster residues. The combined data indicate that Arg 23 is a major determinant for interfacial phospholipid binding and participates in an intermolecular salt bridge that is key for trimer formation on the membrane surface. Together, crystallographic and solution data provide evidence that the interfacial basic cluster is a locus where trimerization is synergistically coupled to membrane phospholipid binding.Annexins comprise a large family of homologous eukaryotic proteins with a common core region that promotes Ca 2ϩ -dependent binding to phospholipid membrane surfaces (for a review, see Refs. 1-5). This property underlies many proposed in vivo annexin functions, including membrane trafficking, cell signaling, and roles in inflammatory and coagulation processes. Annexin V is an abundant protein that binds preferentially to acidic phospholipid membranes in the presence of Ca 2ϩ . The annexin-phospholipid association is of high affinity but is reversible with the removal of Ca 2ϩ . When bound to the membrane surface, annexin V molecules assemble laterally into well organized trimers and higher order arrays of trimers (6). This lateral assembly promotes spontaneous two-dimensional crystallization of annexin V on membrane surfaces, a process that is inhibited by extreme membrane surface curvature (7,8). Annexin-coated membrane surfaces undergo changes in their properties, becoming markedly rigid as the crystallization domains increase in size (9 -11). This membrane-bound layer of annexin V, which has anticoagulant properties, is believed to play a functional role in processes associated with the protein. In placenta, reduced levels of annexin V occur with anti-phospholipid syndrome, which causes hypercoagulation and recurrent miscarriage. Rand and co-workers (12, 13) have proposed a mechanism for anti-phospholipid syndrome in which the disease process strips off the protective layer of annexin V at the maternal-fetal interface, permitting excessive thrombosis to occur.The mechanism of membrane adsorption of annexin V has generated considerable interest from many disciplines, from clinical to structural biology. The protein provides an excellent model for elucidating complex interfacial behavior by peripheral membrane proteins. The molecular structure of the 35-kDa protein has been well characterized in both soluble and membrane-bound forms by x-ray ...

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Section: Promotion Of Subunit Interactions By Basic Clustermentioning

confidence: 99%

mentioning

confidence: 99%

Interfacial Basic Cluster in Annexin V Couples Phospholipid Binding and Trimer Formation on Membrane Surfaces

Mo¹,

Campos²,

Mealy³

et al. 2003

Journal of Biological Chemistry

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“…Oling et al, 2001;Parcej and Eckhardt-Strelau, 2003;Tahara et al, 2000;Ziegler et al, 2004). In those cases, a single particle approach can lead to a substantial improvement (Tahara et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

A maximum likelihood approach to two-dimensional crystals

Zeng

Stahlberg

Grigorieff

2007

Journal of Structural Biology

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Maximum Likelihood (ML) processing of transmission electron microscopy images of protein particles can produce reconstructions of superior resolution due to a reduced reference bias. We have investigated a ML processing approach to images centered on the unit cells of two-dimensional (2D) crystal images. The implemented software makes use of the predictive lattice node tracking in the MRC software, which is used to window particle stacks. These are then noise-whitened and subjected to ML processing. Resulting ML maps are translated into amplitudes and phases for further processing within the 2dx software package. Compared with ML processing for randomly oriented single particles, the required computational costs are greatly reduced as the 2D crystals restrict the parameter search space. The software was applied to images of negatively stained or frozen hydrated 2D crystals of different crystal order. We find that the ML algorithm is not free from reference bias, even though its sensitivity to noise correlation is lower than for pure cross-correlation alignment. Compared with crystallographic processing, the newly developed software yields better resolution for 2D crystal images of lower crystal quality, and it performs equally well for well-ordered crystal images.

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“…ANX A5 molecules are monomeric in solution, however, after binding to phospholipid membrane, they spontaneously form tightly bound trimers. The membrane bound trimers self-organize into two types of 2D crystals, with p3 or p6 symmetry (Oling et al, 1998). The mechanism of ANX A5 crystallization on negatively charged phospholipid membranes in the presence of calcium has been thoroughly described (Reviakine et al, 1998(Reviakine et al, , 2001Irman et al, 2009) and it is here schematically shown in Figure 4.…”

Section: Annexin A5's Characteristicsmentioning

confidence: 99%

Protein Interactions on Phospholipid Bilayer, Studied by AFM Under Physiological Conditions

Irman¹,

Škarabot²,

Muševič³

et al. 2012

Atomic Force Microscopy Investigations Into Biology - From Cell to Protein

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Trimers, Dimers of Trimers, and Trimers of Trimers Are Common Building Blocks of Annexin A5 Two-Dimensional Crystals

Cited by 111 publications

References 26 publications

Interfacial Basic Cluster in Annexin V Couples Phospholipid Binding and Trimer Formation on Membrane Surfaces

Interfacial Basic Cluster in Annexin V Couples Phospholipid Binding and Trimer Formation on Membrane Surfaces

A maximum likelihood approach to two-dimensional crystals

Protein Interactions on Phospholipid Bilayer, Studied by AFM Under Physiological Conditions

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