Preparation and Characterization of Posttranslationally Modified Tubulins From Artemia franciscana
Bibliographic record
Abstract
Tubulin heterogeneity within eukaryotic cells is generated by differential gene expression and posttranslational modification of alpha- and beta-tubulin gene products, either as heterodimers or when polymerized into microtubules. The characterization of posttranslationally modified tubulins from the crustacean Artemia franciscana is presented, although tubulins from other sources can be studied with these procedures. Tubulin is prepared from cell free extracts by taxol-induced assembly and centrifugation of microtubules through sucrose cushions, which also yields microtubule-associated proteins, or it is purified to apparent homogeneity by relatively simple chromatographic procedures and assembly/disassembly steps. To detect posttranslationally modified tubulins protein samples are electrophoresed in sodium dodecyl sulfate (SDS) polyacrylamide gels, blotted to nitrocellulose membranes and probed with isoform-specific antibodies. Isotubulins, for which gene-encoded amino acid differences and posttranslational modifications generate charge variations, are resolved in two-dimensional gels using isoelectric focusing followed by SDS polyacrylamide gel electrophoresis, a procedure useful for resolution of microtubule-associated proteins. Isoforms patterns are visualized by Coomassie blue and/or silver staining and individual isoforms are identified by antibody reactivity on Western blots. Tubulin isoforms are localized in Artemia by immunofluorescent staining of larvae. The focus of this chapter is the purification of tubulin from a nonneural source and characterization of tyrosinated, detyrosinated, and nontyrosinatable alpha-tubulins using polyclonal antibodies made to carboxy-terminal peptides of each isoform.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".