Reagents for Rapid Reduction of Native Disulfide Bonds in Proteins

Singh, Rajeeva; Whitesides, G. M.

doi:10.1006/bioo.1994.1008

Cited by 22 publications

(13 citation statements)

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“…We used DTT, a common reductive reagent known to break disulfide bonds in proteins, to further react with the extracted keratins to synthesize reduced Keratin‐SH at room temperature (Figure a). The free thiol content of the reduced Keratin‐SH samples was determined by the Ellman's assay as ∼0.55 mmol/g, which was significantly higher than the pristine keratin samples.…”

Section: Resultsmentioning

confidence: 99%

Visible light crosslinkable human hair keratin hydrogels

Yue

Liu

Byambaa

et al. 2018

Bioengineering & Transla Med

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Keratins extracted from human hair have emerged as a promising biomaterial for various biomedical applications, partly due to their wide availability, low cost, minimal immune response, and the potential to engineer autologous tissue constructs. However, the fabrication of keratin‐based scaffolds typically relies on limited crosslinking mechanisms, such as via physical interactions or disulfide bond formation, which are time‐consuming and result in relatively poor mechanical strength and stability. Here, we report the preparation of photocrosslinkable keratin‐polyethylene glycol (PEG) hydrogels via the thiol‐norbornene “click” reaction, which can be formed within one minute upon irradiation of visible light. The resulting keratin‐PEG hydrogels showed highly tunable mechanical properties of up to 45 kPa in compressive modulus, and long‐term stability in buffer solutions and cell culture media. These keratin‐based hydrogels were tested as cell culture substrates in both two‐dimensional surface seeding and three‐dimensional cell encapsulation, demonstrating excellent cytocompatibility to support the attachment, spreading, and proliferation of fibroblast cells. Moreover, the photocrosslinking mechanism makes keratin‐based hydrogel suitable for various microfabrication techniques, such as micropatterning and wet spinning, to fabricate cell‐laden tissue constructs with different architectures. We believe that the unique features of this photocrosslinkable human hair keratin hydrogel promise new opportunities for their future biomedical applications.

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Section: Resultsmentioning

confidence: 99%

Visible light crosslinkable human hair keratin hydrogels

Yue

Liu

Byambaa

et al. 2018

Bioengineering & Transla Med

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“…However DTT, as well as other thiols, is very ineffective at low pH and totally inactive at pH 1.5 (Han & Han, 1994). Since DTT has a high pKa value thiol (9.2), at pH 7.0 or lower only a small fraction of its thiol groups are present in the thiolate reactive form (Singh & Whitesides, 1994) and its action will be slow to none at acidic pH. Under these conditions, the predominant species will be the protonated non-reactive thiol.…”

Section: Interaction Of So 2 With Protein Fractions Differing In Hydrmentioning

confidence: 99%

The challenging SO2-mediated chemical build-up of protein aggregates in wines

et al. 2016

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“…The reactivity of a dithiol depends on the pK a value. In the reduction of disulfide bonds, the maximum apparent rate of a thiol-disulfide interchange is observed for a thiol whose pK a is approximately equal to the pH of the solution [53, 54]. The pH of the fertilization medium is approximately 7.8, which is lower than the pK a of DTT (9.0), whereas the thiol pK a value of DTBA is approximately 8.2.…”

Section: Discussionmentioning

confidence: 99%

Pretreatment of bovine sperm with dithiobutylamine (DTBA) significantly improves embryo development after ICSI

Suttirojpattana

Somfai

Matoba

et al. 2016

Journal of Reproduction and Development

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We assessed the effect of pretreating sperm with dithiobutylamine (DTBA) to improve embryo development by intracytoplasmic sperm injection (ICSI) in cows. Acridine Orange staining revealed that when applied at different concentrations (2.5, 5, and 10 mM) and exposure times (5 min, 20 min, 1 h, and 2 h), DTBA reduced disulfide bonds in spermatozoa with the highest efficacy at 5 mM for 5 min. DTBA enhanced the percentage of spermatozoa with free protamine thiol groups compared with untreated spermatozoa (control) (P < 0.05); however, this result did not differ from that of dithiothreitol (DTT) treatment. The percentage of live spermatozoa after DTBA treatment was identical to that in the control, but significantly higher than that after DTT treatment (P < 0.05). After ICSI, DTBA treatment tended to improve male pronuclear formation rate (P = 0.071) compared with non-treated sperm injection. Blastocyst formation rate was significantly improved by DTBA treatment compared with that in DTT, control, and sham injection groups (P < 0.05). Blastocyst quality in terms of cell numbers and ploidy was not different among these groups. In conclusion, DTBA increases the efficacy of blastocyst production by ICSI even if DTT treatment does not work.

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Reagents for Rapid Reduction of Native Disulfide Bonds in Proteins

Cited by 22 publications

References 0 publications

Visible light crosslinkable human hair keratin hydrogels

Visible light crosslinkable human hair keratin hydrogels

The challenging SO2-mediated chemical build-up of protein aggregates in wines

Pretreatment of bovine sperm with dithiobutylamine (DTBA) significantly improves embryo development after ICSI

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