Evolutionary Divergence of Substrate Specificity within the Chymotrypsin-like Serine Protease Fold

Perona, John J.; Craik, Charles S.

doi:10.1074/jbc.272.48.29987

Cited by 328 publications

(277 citation statements)

References 36 publications

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“…Moreover, it contains six cysteine residues typically present in invertebrate serine proteases (Fig.·2) (Yan et al, 2001). The presence of the three amino acids glycine, glycine and aspartate in the primary specificity pocket suggests that CTLP1 may exhibit a chymotrypsin-like substrate specificity with a glycine at the bottom of the pocket as the primary determinant (Perona and Craik, 1997). However, the occurrence of an aspartate residue at the side of the pocket may alter the specificity of CTLP1 significantly.…”

Section: Isolation and Sequencing Of The Cdna Encoding Ctlp1mentioning

confidence: 99%

A chymotrypsin-like serine protease interacts with the chitin synthase from the midgut of the tobacco hornworm

Broehan

Zimoch

Wessels

et al. 2007

Journal of Experimental Biology

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The chitin portion of the peritrophic matrix in the midgut of the tobacco hornworm, Manduca sexta, is produced by chitin synthase 2 (CHS2), a transmembrane family II glycosyltransferase, located at the apical tips of brush border microvilli. To look for proteins that potentially interact with CHS2, we performed yeast twohybrid screening, identifying a novel chymotrypsin-like protease (CTLP1) that binds to the extracellular carboxyterminal domain of CHS2. The occurrence of this interaction in vivo is supported by co-localization and coimmunoprecipitation data. Based on our findings we propose that chitin synthesis is controlled by an intestinal proteolytic signalling cascade linking chitin synthase activity to the nutritional state of the larvae.

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Section: Isolation and Sequencing Of The Cdna Encoding Ctlp1mentioning

confidence: 99%

A chymotrypsin-like serine protease interacts with the chitin synthase from the midgut of the tobacco hornworm

Broehan

Zimoch

Wessels

et al. 2007

Journal of Experimental Biology

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“…Students are then informed that chymotrypsin residues 189, 190, and 228 are three of the residues lining the specificity pocket of the enzyme [12]. They are asked to name the residues at these positions for chymotrypsin and determine the corresponding residues for trypsin, based on their sequence alignment.…”

Section: Students Are Given Detailed Instructions For Obtain-mentioning

confidence: 99%

“…Although other features of these two enzymes are essential for their differing specificities, the residues lining the specificity pocket make an important contribution to their specificities [12].…”

Section: Students Are Given Detailed Instructions For Obtain-mentioning

confidence: 99%

Introductory bioinformatics exercises utilizing hemoglobin and chymotrypsin to reinforce the protein sequence‐structure–function relationship

Inlow¹,

Miller²,

Pittman³

2007

Biochem Molecular Bio Educ

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We describe two bioinformatics exercises intended for use in a computer laboratory setting in an upperlevel undergraduate biochemistry course. To introduce students to bioinformatics, the exercises incorporate several commonly used bioinformatics tools, including BLAST, that are freely available online. The exercises build upon the students' background knowledge of hemoglobin and chymotrypsin, and foster a better understanding of how protein sequence relates to structure and function.

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“…It is known for serine proteases that amino acid residues within the active-site architecture that interact directly with the substrate define the substrate specificity (18,19). Because of these specific interactions, it is possible to alter substrate specificity for serine proteases in general (20,21), and APC in particular (22,23), by substitution of amino acids that form direct, energetically important contacts with the various substrate side chains.…”

mentioning

confidence: 99%

Engineering the proteolytic specificity of activated protein C improves its pharmacological properties

Berg

Gerlitz²,

Shang³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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Human activated protein C (APC) is an antithrombotic, antiinflammatory serine protease that plays a central role in vascular homeostasis, and activated recombinant protein C, drotrecogin alfa (activated), has been shown to reduce mortality in patients with severe sepsis. Similar to other serine proteases, functional APC levels are regulated by the serine protease inhibitor family of proteins including ␣1-antitrypsin and protein C inhibitor. Using APC-substrate modeling, we designed and produced a number of derivatives with the goal of altering the proteolytic specificity of APC such that the variants exhibited resistance to inactivation by protein C inhibitor and ␣1-antitrypsin yet maintained their primary anticoagulant activity. Substitutions at Leu-194 were of particular interest, because they exhibited 4-to 6-fold reductions in the rate of inactivation in human plasma and substantially increased pharmacokinetic profiles compared with wild-type APC. This was achieved with minimal impairment of the anticoagulant͞ antithrombotic activity of APC. These data demonstrate the ability to selectively modulate substrate specificity and subsequently affect in vivo performance and suggest therapeutic opportunities for the use of protein C derivatives in disease states with elevated serine protease inhibitor levels.

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Evolutionary Divergence of Substrate Specificity within the Chymotrypsin-like Serine Protease Fold

Cited by 328 publications

References 36 publications

A chymotrypsin-like serine protease interacts with the chitin synthase from the midgut of the tobacco hornworm

A chymotrypsin-like serine protease interacts with the chitin synthase from the midgut of the tobacco hornworm

Introductory bioinformatics exercises utilizing hemoglobin and chymotrypsin to reinforce the protein sequence‐structure–function relationship

Engineering the proteolytic specificity of activated protein C improves its pharmacological properties

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