Structure of the iron‐free true C‐terminal half of bovine lactoferrin produced by tryptic digestion and its functional significance in the gut

Rastogi, Nilisha; Singh, Avinash; Pandey, Sada N.; Sinha, Mau; Bhushan, A.; Kaur, Punit; Sharma, Sujata

doi:10.1111/febs.12827

Cited by 14 publications

(27 citation statements)

References 47 publications

(70 reference statements)

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“…Interestingly, the C-terminal lobe has been recently reported to be resistant to trypsin digestion and retains antimicrobial activity through the conservation of a defined ironbinding site (Rastogi et al, 2014a;Rastogi et al, 2014b), which may be important when considering use of oral lactoferrin supplements and transit through the gastrointestinal tract.…”

Section: Lactoferrinmentioning

confidence: 99%

A role for whey-derived lactoferrin and immunoglobulins in the attenuation of obesity-related inflammation and disease

Brimelow

West

Williams

et al. 2015

Critical Reviews in Food Science and Nutrition

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Obesity is a strong predictive factor in the development of chronic disease and has now superseded undernutrition as a major public health issue. Chronic inflammation is one mechanism thought to link excess body weight with disease. Increasingly, the gut and its extensive population of commensal microflora are recognized as playing an important role in the development of obesity-related chronic inflammation. Obesity and a high fat diet are associated with altered commensal microbial communities and increased intestinal permeability which contributes to systemic inflammation as a result of the translocation of lipopolysaccharide into the circulation and metabolic endotoxemia. Various milk proteins are showing promise in the prevention and treatment of obesity and chronic low-grade inflammation via reductions in visceral fat, neutralization of bacteria at the mucosa and reduced intestinal permeability. In this review, we focus on evidence supporting the potential antiobesogenic and anti-inflammatory effects of bovine whey-derived lactoferrin and immunoglobulins.

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Section: Lactoferrinmentioning

confidence: 99%

A role for whey-derived lactoferrin and immunoglobulins in the attenuation of obesity-related inflammation and disease

Brimelow

West

Williams

et al. 2015

Critical Reviews in Food Science and Nutrition

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“…The previously reported procedure was used for the purification of lactoferrin . The preparation of C‐lobe was carried out using limited proteolysis with trypsin as described earlier . The purification of C‐lobe was also adopted from the previous studies …”

Section: Methodsmentioning

confidence: 99%

“…Although C‐lobe was also cloned and expressed but no structural studies were reported. Later on, C‐lobe was successfully produced by limited proteolysis using serine proteases . The first structure of a not so perfect C‐lobe consisting of residues, Tyr342–Ser676 and Leu681–Ala685 of bovine lactoferrin (BLF) (residues, Ala1–Arg689) produced by hydrolyzing lactoferrin using proteinase K (PK‐C‐lobe), was determined in iron saturated form (Holo‐PK‐C‐lobe) .…”

Section: Introductionmentioning

confidence: 99%

“…The first structure of a not so perfect C‐lobe consisting of residues, Tyr342–Ser676 and Leu681–Ala685 of bovine lactoferrin (BLF) (residues, Ala1–Arg689) produced by hydrolyzing lactoferrin using proteinase K (PK‐C‐lobe), was determined in iron saturated form (Holo‐PK‐C‐lobe) . The second structure of C‐lobe, a perfect C‐lobe consisting of residues, Tyr342–Arg689, produced by hydrolyzing lactoferrin using trypsin (TN‐C‐lobe), was determined in the iron‐free form (Apo‐TN‐C‐lobe) . Since C‐lobe produced with tryptic digestion is perfect in size as well as it is physiologically more relevant than that produced using proteinase K, the structure determinations of such a perfect C‐lobe in the iron saturated form at various physiologically relevant pH will provide valuable insights into the iron binding and release.…”

Section: Introductionmentioning

confidence: 99%

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Structure of iron saturated C‐lobe of bovine lactoferrin at pH 6.8 indicates a weakening of iron coordination

Rastogi

Singh

et al. 2016

Proteins

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The bilobal lactoferrin is an approximately 76 kDa glycoprotein. It sequesters two Fe(3+) ions together with two CO(3)(2-) ions. The C-terminal half (residues, Tyr342-Arg689, C-lobe) of bovine lactoferrin (BLF) (residues Ala1-Arg689) was prepared by limited proteolysis using trypsin. Both C-lobe and intact BLF were saturated to 100%. Both of them retained up to nearly 85% of iron at pH 6.5. At pH 5.0, C-lobe retained 75% of iron whereas intact protein could retain only slightly more than 60%. At pH 4.0 both contained 25% iron and at pH 2.0 they were left with iron concentration of only 10%. The structure of iron saturated C-lobe was determined at 2.79 Å resolution and refined to R(cryst) and R(free) factors of 0.205 and 0.273, respectively. The structure contains two crystallographically independent molecules, A and B. They were found to have identical structures with an r.m.s. shift of 0.5 Å for their C(α) atoms. A high solvent content of 66% was observed in the crystals. The average value of an overall B-factor was 68.0 Å(2). The distance of 2.9 Å observed for the coordination bond between Fe(3+) ion and N(e2) of His595 appeared to be considerably longer than the normally observed values of 1.9-2.2 Å. This indicated that the coordination bond involving His595 may be absent. Other coordination distances were observed in the range of 2.1-2.3 Å. Based on the present structure of iron saturated C-lobe, it may be stated that His595 is the first residue to dissociate from ferric ion when the pH is lowered.

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“…That is an extensive contribution. (Salunke et al, 1985;Banerjee et al, 1994Banerjee et al, , 1996Sankaranarayanan et al, 1996;Singh et al, 2001;Sharma et al, 2011;Rastogi et al, 2014) We need to include many areas like non-aqueous enzymology, enzyme promiscuity and moonlighting proteins in our syllabi and encourage people to work in these areas. The first one offers an opportunity for biocatalyst-based synthesis of industrial and drug intermediates, agrochemicals and biosurfactants (Gupta, 1992;Halling, 1994;Carrea and Riva, 2000;Hudson et al, 2005).…”

Section: Applied Bio-catalysis and "Make In India" Programmentioning

confidence: 99%

Skills in biocatalysis are essential for establishing a sound biotechnological base in India

Gupta¹

2015

Proceedings of the Indian National Science Academy

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Techniques developed by enzymologists and protein chemists have been indispensible in many areas of biotechnology. Bio-separation of proteins, biocatalyst engineering and medium engineering are important examples of this. Our understanding of enzyme specificity has helped us in gaining insight into catalytic promiscuity. This, along with the possibility of carrying out biocatalysis in organic solvents has widened the scope of using enzymes for organic synthesis.Reestablishing a strong base in enzymology is crucial towards creating a strong base for the field of biotechnology in India. Keywords IntroductionQuite a few aspects of biotechnology involve production and applications of enzymes. In the old days, most of these aspects were covered under industrial enzymology (Godfrey and Reichelt, 1983). Gradually, a more broad based term 'applied biocatalysis' came into vogue (Straathof and Adlercreutz, 2000) which was an interface between enzymology and biochemical engineering. This phase was characterized by few new developments: (1) As a result of the rDNA technology, our capability to produce mutant properties grew exponentially. There was a scramble to produce proteins at a lower cost. The stakes were no longer limited to low cost industrial grade enzymes. Costlier proteins used in molecular biology, diagnostics and biosensors and pharmaceutical proteins/enzymes could be produced by cloning and their properties tailored to suit the applications. The result was a paradigm shift in the downstream processing methods. Shorter and efficient protocols emerged. Affinity chromatography, rather than being a 'polishing' step at the end, was promoted to the earlier steps of purification. In fact, upstream and downstream processing steps were integrated (Gupta, 2002;Przybycien et al., 2004;Mondal et al., 2006). (2) It was realized that enzymes could be used in non aqueous media (Carrea and Riva 2000; Gupta 2000;Patel 2000; Vulfson et al., 2001). This opened up novel applications; especially in the synthesis of chiral compounds. Table 1 lists some of the milestones in the area of applied biocatalysis.It is remarkable how much enzymology contributed to all this.The present review attempts to emphasize the importance of the skills which are part of the toolbox Published Online on 10 December 2015 1114Munishwar Nath Gupta and Joyeeta Mukherjee et al., 1973; Mosbach 1976 Mosbach , 1980 Nilsson and Mosbach 1981; Mattiasson 1983;Mattiasson 1988) Berezin's group in Moscow was the third one (Berezin 1978; Mozhaev et al., 1987;Martinek and Mozhaev 1993). Current work is adequately discussed at many places ( Enzyme deactivation models Most people wrongly give half lives without analysing the kinetics of deactivation. Sadana's work needs to be known more widely. (Sadana 1991(Sadana , 1993 Enzymes as reverse micelles This is another approach in low water enzymology. (Luisi 1985; Larsson et al., 1987; Adlercreutz et al., 1988) Enzyme use in biosensors Immobilization allows enzymes to interface with transducers. (Yarapolov et al...

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Structure of the iron‐free true C‐terminal half of bovine lactoferrin produced by tryptic digestion and its functional significance in the gut

Cited by 14 publications

References 47 publications

A role for whey-derived lactoferrin and immunoglobulins in the attenuation of obesity-related inflammation and disease

A role for whey-derived lactoferrin and immunoglobulins in the attenuation of obesity-related inflammation and disease

Structure of iron saturated C‐lobe of bovine lactoferrin at pH 6.8 indicates a weakening of iron coordination

Skills in biocatalysis are essential for establishing a sound biotechnological base in India

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