Isolation and partial characterization of cytoplasmic α agglutination substance in the yeast <i>Saccharomyces cerevisiae</i>

Yamaguchi, Makoto; Yoshida, Kazuo; Yanagishima, Naohiko

doi:10.1016/0014-5793(82)80502-4

Cited by 15 publications

(15 citation statements)

References 14 publications

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“…In equilibrium binding experiments with radioactively labeled ␣-agglutinin, low temperature had little effect on the binding isotherm. However, previous studies showed that low temperature does reduce cellular agglutinability both in vivo and in vitro (41,43). SPR was used to obtain the k off and the k on and to detect the effect of low temperature on the interaction of the agglutinins in real time.…”

Section: ϫ10mentioning

confidence: 99%

Interaction of α-Agglutinin and a -Agglutinin, Saccharomyces cerevisiae Sexual Cell Adhesion Molecules

et al. 2001

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␣-Agglutinin and a-agglutinin are complementary cell adhesion glycoproteins active during mating in the yeast Saccharomyces cerevisiae. They bind with high affinity and high specificity: cells of opposite mating types are irreversibly bound by a few pairs of agglutinins. Equilibrium and surface plasmon resonance kinetic analyses showed that the purified binding region of ␣-agglutinin interacted similarly with purified a-agglutinin and with a-agglutinin expressed on cell surfaces. At 20°C, the K D for the interaction was 2 ؋ 10 ؊9 to 5 ؋ 10 ؊9M. This high affinity was a result of a very low dissociation rate (Ϸ 2.6 ؋ 10 ؊4 s ؊1 ) coupled with a low association rate ‫؍(‬ 5 ؋ 10 4 M ؊1 s ؊1 ). Circular-dichroism spectroscopy showed that binding of the proteins was accompanied by measurable changes in secondary structure. Furthermore, when binding was assessed at 10°C, the association kinetics were sigmoidal, with a very low initial rate. An induced-fit model of binding with substantial apposition of hydrophobic surfaces on the two ligands can explain the observed affinity, kinetics, and specificity and the conformational effects of the binding reaction.

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Section: ϫ10mentioning

confidence: 99%

Interaction of α-Agglutinin and a -Agglutinin, Saccharomyces cerevisiae Sexual Cell Adhesion Molecules

et al. 2001

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“…The a-and a-agglutinins are large glycoproteins, with both N-linked and 0-linked carbohydrates (45,55,56; P. N. Lipke, unpublished results). Other yeast species express analogous agglutinins (7,28,33,54).…”

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confidence: 95%

AG alpha 1 is the structural gene for the Saccharomyces cerevisiae alpha-agglutinin, a cell surface glycoprotein involved in cell-cell interactions during mating.

Lipke¹,

Wojciechowicz²,

Kurjan³

1989

Mol. Cell. Biol.

151

143

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We have cloned the a-agglutinin structural gene, AGal, by the isolation of a-specific agglutination-defective mutants, followed by isolation of a complementing plasmid. Independently isolated a-specific agglutinationdefective mutations were in a single complementation group, consistent with biochemical results indicating that the a-agglutinin is composed of a single polypeptide. Mapping results suggested that the complementation group identified by these mutants is allelic to the agal mutation identified previously. Expression ofAGal RNA was a specific and inducible by a-factor. Sequences similar to the consensus sequences for positive control by MATal and pheromone induction were found upstream of the AGaJ initiation codon. The AGal gene could encode a 650-amino-acid protein with a putative signal sequence, 12 possible N-glycosylation sites, and a high proportion of serine and threonine residues, all of which are features expected for the a-agglutinin sequence. cells of other yeast species (7,54). Therefore, the glycoprotein-glycoprotein interactions are quite specific even though the analogous agglutinins of different yeast species share biochemical characteristics.Biochemical analysis has indicated that the a-agglutinin of S. cerevisiae is a glycoprotein of approximate 160 kilodaltons (kDa) (45). Removal of N-linked carbohydrate generates a major species of 72 kDa and several minor species. Structural analysis and determination of agglutinin binding determinants have been hampered by low yields and carbohydrate-generated heterogeneity of the agglutinins. A genetic approach allowed the isolation of a putative a-agglutinin structural mutant, agal (43). The agal mutation results in an a-specific agglutination defect, resulting from the lack of expression of active a-agglutinin, and a decrease in a mating efficiency. Additional mutants have been isolated that result in agglutination defects in both a and a cells and also affect pheromone production and response (17); the pleiotropic effects of these mutations indicate that they do not identify agglutinin structural genes.We have initiated a genetic and molecular approach to the study of agglutinin structure and function, which will complement the biochemical approach that has been used previ-

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“…In the genus Saccharomyces, adhesive components from both mating types of Saccharomyces kluyveri and of Saccharomyces cerevisiae have been isolated (16,22,27,30,31). In all cases, cell-adhesive components appear to be monovalent.…”

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Interaction of alpha-agglutinin with Saccharomyces cerevisiae a cells

Lipke¹,

Terrance²,

Wu³

1987

J Bacteriol

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Binding of Saccharomyces cerevisiae alpha-agglutinin to target a cells was assayed by agglutination inhibition and 125I-alpha-agglutinin binding. The assays showed characteristics of equilibrium binding, namely saturability, competability, and the establishment of a kinetic endpoint in the presence of free alpha-agglutinin and free receptor. The binding was heterogeneous, displaying strong binding (10(9) liters/mol) and a weaker interaction. There were about 2 X 10(4) strong binding sites per a cell. Denaturing gels displayed identical labeled species binding to the a cells in the weak and strong interactions. Furthermore, weakly bound material could subsequently bind tightly to fresh a cells, implying that the same species of alpha-agglutinin was bound in the two states.

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Isolation and partial characterization of cytoplasmic α agglutination substance in the yeast Saccharomyces cerevisiae

Cited by 15 publications

References 14 publications

Interaction of α-Agglutinin and a -Agglutinin, Saccharomyces cerevisiae Sexual Cell Adhesion Molecules

Interaction of α-Agglutinin and a -Agglutinin, Saccharomyces cerevisiae Sexual Cell Adhesion Molecules

AG alpha 1 is the structural gene for the Saccharomyces cerevisiae alpha-agglutinin, a cell surface glycoprotein involved in cell-cell interactions during mating.

Interaction of alpha-agglutinin with Saccharomyces cerevisiae a cells

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