Amyloidogenicity of recombinant human pro-islet amyloid polypeptide (ProIAPP).

BACKGROUND: Pancreatic amyloid has been associated with type II diabetes. The major constituent of pancreatic amyloid is the 37-residue peptide islet amyloid polypeptide (IAPP). IAPP is expressed as a 67-residue pro-peptide called ProIAPP which is processed to IAPP following stimulation. While the molecular events underlying IAPP amyloid formation in vitro have been studied, little is known about the role of ProIAPP in the formation of pancreatic amyloid. This has been due in part to the limited availability of purified ProIAPP for conformational and biochemical studies. RESULTS: We present a method for efficient recombinant expression and purification of ProIAPP and a processing site mutant, mutProIAPP, as thioredoxin (Trx) fusion proteins. Conformation and amyloidogenicity of cleaved ProIAPP and mutProIAPP and the fusion proteins were assessed by circular dichroism, electron microscopy and Congo red staining. We find that ProIAPP and mutProIAPP exhibit strong self-association potentials and are capable of forming amyloid. However, the conformational transitions of ProIAPP and mutProIAPP during aging and amyloidogenesis are distinct from the random coil-to-beta-sheet transition of IAPP. Both proteins are found to be less amyloidogenic than IAPP and besides fibrils a number of non-fibrillar but ordered aggregates form during aging of ProIAPP. ProIAPP aggregates are cytotoxic on pancreatic cells but less cytotoxic than IAPP while mutProIAPP aggregates essentially lack cytotoxicity. The Trx fusion proteins are neither amyloidogenic nor cytotoxic. CONCLUSIONS: Our studies suggest that ProIAPP has typical properties of an amyloidogenic polypeptide but also indicate that the pro-region suppresses the amyloidogenic and cytotoxic potentials of IAPP.

Argument structure frames: a lexical complexity metric?

The number of semantic argument structure frames associated with a verb has been reported to influence ease of processing during language comprehension. The present experiments tested the generality of the argument structure complexity effect with three dependent measures: eye-fixation times, naming latencies, and lexical decision latencies. Two eye-movement experiments and two experiments using cross-modal tasks failed to provide evidence supporting the argument structure complexity effect. The present experiments indicated that results reflecting verbs' argument structure complexity are not generalizable.

Stress grouping improves performance on an immediate serial list recall task.

Five experiments investigated whether perceptual patterning afforded by imposing a recurrent stress pattern on auditorially presented lists has a positive effect on list recall. The experiments also addressed whether the recall advantage reflected the salience that the stress pattern created for certain items or whether the recall advantage arose from the distinct grouping configurations that were produced by the stress pattern. The authors explored these issues by examining immediate serial-recall performance for spoken lists that either did or did not have a stress pattern imposed on them. Lists had an anapest or dactylic stress pattern or were monotone and consisted of two stimulus types, either digit names or common English nouns. Results showed that stress patterns enhanced serial-recall performance and that the recall benefit derived primarily from the perceptual grouping afforded by the stress patterns. Results also showed that the grouping benefit derived from stress patterning generalizes to monotone lists. In contrast, salience effects are attached to the stimulus per se and do not transfer.

On the conditions necessary for obtaining argument structure complexity effects.

Schmauder (1991), studying eye movements during reading, cross-modal naming, and cross-modal lexical decision (CMLD) tasks, failed to find evidence of verb argument structure complexity as Shapiro, Zurif, and Grimshaw (1987) had reported for the CMLD task. Shapiro, Brookins, Gordon, and Nagel (1991) suggested that Schmauder did not detect the effect in the CMLD task because the monosyllabic secondary lexical decision (LD) probes she used did not produce enough processing load to detect an effect of argument structure complexity. The present experiment compared the LD probes used by Schmauder with the LD probes used by Shapiro et al. (1987) and failed to find any evidence for the argument structure complexity effect for either type of probe.

Ability to stand alone and processing of open-class and closed-class words: isolation versus context.

Four experiments investigating processing of closed-class and open-class words in isolation and in sentence contexts are reported. Taft (1990) reported that closed-class words which could not meaningfully stand alone and open-class words which could not meaningfully stand alone incurred longer lexical decision responses than did control words. Taft also reported that closed-class and open-class words which could stand alone meaningfully were not associated with longer lexical decision responses than were control words. Experiments 1 and 2 replicated Taft's effect of ability to stand alone on lexical decision responses to closed-class and open-class words presented in isolation. In Experiments 3 and 4, the same lexical decision targets were presented as part of semantically neutral context sentences in a moving window paradigm. The stand-alone effect was not present in Experiments 3 and 4. The results suggest Taft's conclusion that meaningfulness of a word influences lexical decision needs revision. An explanation is provided according to which support from message level and syntactic and lexical sources in sentence contexts influence words' perceived "meaningfulness."

The influence of semantic fit on on-line sentence processing.

The interactive influence of verb complement preferences and noun phrase semantic fit on resolution of temporary syntactic ambiguity was investigated in an eye movement experiment. The present semantic fit manipulation included noun phrases that fit well as direct objects of the verbs that they followed and noun phrases that were possible but less likely direct objects of the verbs in question. This contrasted with existing research on the use of verb complement preferences and semantic fit during sentence processing, in which processing of noun phrases that are possible direct objects has been compared with processing of noun phrases that are not possible direct objects of the verbs that they follow. Verb complement preference information and noun phrase semantic fit interacted at early stages of on-line sentence processing. Implications of these results for interactive and structural models of sentence processing are discussed.

Lexical processing and text integration of function and content words: evidence from priming and eye fixations.

The results of two experiments comparing processing of function words and content words are reported. In Experiment 1, priming was present for both related function and related content word pairs, as measured in lexical decision response times. In Experiment 2, participants' eye movements were monitored as they read sentences containing either a high- or a low-frequency function or content target word. Average word length and word frequency were matched across the function and content word conditions. Function words showed frequency effects in first-fixation and gaze duration that were similar to those seen for content words. Clear differences in on-line processing of function and content words emerged in later processing measures. These differences were reflected in reading patterns and reading time measures. There was inflated processing time in the phrase immediately following a low-frequency function word, and participants made more regressions to the target word in this condition than in the other three conditions. The priming effects in lexical decision and the word frequency effects in initial processing measures in silent reading for both word types were taken as evidence of common lexical processing for function and content words. The observed differences in later processing measures in the eye-movement data were taken as evidence of differences in the role that the two word types have in sentence processing beyond the lexical level.