Skip to content
EvidenceHelixEvidenceLib · Scientific Encyclopedia
⌘ K

Scientific library

EvidenceLib encyclopediaAll articles A–Z2,100Open textbooks2Cancer study catalog458How it is made

My workspace this browser

Saved pages0Notes & annotations0

Recently viewed

Explore by field

Cancer and Oncology107Genetics and Genomics95Neuroscience110Cardiovascular Medicine105Immunology90Pharmacology99Molecular Biology100Infectious Diseases102Rare and Inherited Diseases106Clinical Research98Anatomy and Physiology542Drugs and Chemicals546
Plans & pricingCompare EvidenceHelix workspace plans
Library›EvidenceLib›Molecular Biology›Substrate Specificity
Explore more with EvidenceHelix Workspace

Compare studies, analyze evidence and create research reports with premium tools.

Explore Researcher plans

Research reading room

Substrate Specificity

Molecular Biology · NLM MeSH · updated 2026-10-29 · version 1

Citations verified against PubMedNot yet clinician-reviewed9 referencesApproximately 3 min read

A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.

Table of Contents

1. Definition2. Research landscape3. Reading the evidence4. Open-access studies5. Key reviews6. References7. About this article

Definition and research guide

This page combines a MeSH definition, publication statistics and selected research abstracts. It is a starting point for reading the literature, not a medical textbook or a systematic evidence review.

Content available: 3 complete abstracts (567 words), 9 references. Full article text is not stored or reproduced here; follow the PMID, DOI or PMC links to read the original sources and their access terms.

Definition

A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.

Definition: NLM Medical Subject Headings (MeSH) scope note (Public domain (U.S. National Library of Medicine); attribution requested). View the MeSH record.

Also known as: Specificities, Substrate; Specificity, Substrate; Substrate Specificities.

Where it fits in the MeSH classification

Broader topics
Biochemical Phenomena

Research landscape

PubMed indexes 103,969 records tagged with the MeSH heading “Substrate Specificity” (counts retrieved 2026-10-29). 8,128 of them (7.8%) were published from 2020 onward, and the busiest period so far is through 1999 with 38,402 records.

Reviews account for 6,895 records (6.6%) and meta-analyses for 11. Randomized controlled trials number 63 (0.1%) as a share of all records on the topic. Randomized trials make up only a small share of this literature, so much of what is known comes from laboratory, observational or descriptive work.

PubMed records by publication periodthrough 1999: 38,402 records; 2000-2009: 31,596 records; 2010-2019: 25,843 records; 2020 onward: 8,128 records38,402through 199931,5962000-200925,8432010-20198,1282020 onward
Records in PubMed indexed with this heading, by publication period. Real counts from NCBI E-utilities; the final period covers only part of the current year. Volume reflects how much has been published, not how strong the evidence is.
Show the data
PeriodRecords
through 199938,402
2000-200931,596
2010-201925,843
2020 onward8,128
Share of records by publication typeReviews: 6.6 percent; Meta-analyses: 0.0 percent; Randomized controlled trials: 0.1 percentReviews6.6% (6,895)Meta-analyses0.0% (11)Randomized controlled trials0.1% (63)
Share of all PubMed records on this heading that carry each publication type. Types overlap with other records, so the shares do not sum to 100%.
Publication-type chart data
Publication types among 103,969 PubMed records
TypeRecordsShare
Reviews6,8956.6%
Meta-analyses110.0%
Randomized controlled trials630.1%

Method: PubMed query "Substrate Specificity"[MeSH Terms] with publication-type and date filters, retrieved 2026-10-29.

Reading the evidence in molecular biology

Molecular biology establishes mechanisms through controlled experiments, often in simplified systems such as purified proteins, cultured cells or model organisms. Such work can show what is possible in a defined system, but whether a pathway matters in a whole organism or in human disease needs further evidence. Check the experimental system, whether findings were reproduced by independent groups, and whether the authors tested the mechanism directly or inferred it from correlation.

Evidence from open-access studies

These open-access studies come from PubMed’s best-match results for this heading (papers whose main topic it is), ranked here by citation count, with one recent addition. Each abstract is reproduced in full under its recorded open licence so that you can read the authors’ own words. They illustrate the literature; they are not a systematic summary of it. Citation counts measure attention, not reliability or clinical benefit.

Study comparison — selected abstracts, not a systematic review
Study / authorsYearSource label / designEurope PMC citationsLicence / source
The intrinsic substrate specificity of the human tyrosine kinome
Yaron-Barir TM, Joughin BA, Huntsman EM, Kerelsky A, Cizin DM, Cohen BM, et al.
2024Research article
Design not recorded in structured metadata
182cc by
PMID 38720073 · DOI · PMC11136658
Targeted protein degradation via intramolecular bivalent glues
Hsia O, Hinterndorfer M, Cowan AD, Iso K, Ishida T, Sundaramoorthy R, et al.
2024Research article
Design not recorded in structured metadata
168cc by
PMID 38383787 · DOI · PMC10917667
Sequence-specific intramembrane proteolysis: identification of a recognition motif in rhomboid substrates
Strisovsky K, Sharpe HJ, Freeman M
2009Research article
Design not recorded in structured metadata
166cc by
PMID 20064469 · DOI · PMC2941825

The intrinsic substrate specificity of the human tyrosine kinome [1]

Research article Yaron-Barir TM, Joughin BA, Huntsman EM, Kerelsky A, Cizin DM, Cohen BM, et al. · · 2024 · cited 182 times (Europe PMC)

Abstract reading summary — source excerpts
Opening excerpt
Phosphorylation of proteins on tyrosine (Tyr) residues evolved in metazoan organisms as a mechanism of coordinating tissue growth1.
Closing excerpt
Finally we show that the intrinsic substrate specificity of Tyr kinases has remained fundamentally unchanged from worms to humans, suggesting that the fidelity between Tyr kinases and their protein substrate sequences has been maintained across hundreds of millions of years of evolution.
Interpretation limits
Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (230 words)

Phosphorylation of proteins on tyrosine (Tyr) residues evolved in metazoan organisms as a mechanism of coordinating tissue growth1. Multicellular eukaryotes typically have more than 50 distinct protein Tyr kinases that catalyse the phosphorylation of thousands of Tyr residues throughout the proteome1-3. How a given Tyr kinase can phosphorylate a specific subset of proteins at unique Tyr sites is only partially understood4-7. Here we used combinatorial peptide arrays to profile the substrate sequence specificity of all human Tyr kinases. Globally, the Tyr kinases demonstrate considerable diversity in optimal patterns of residues surrounding the site of phosphorylation, revealing the functional organization of the human Tyr kinome by substrate motif preference. Using this information, Tyr kinases that are most compatible with phosphorylating any Tyr site can be identified. Analysis of mass spectrometry phosphoproteomic datasets using this compendium of kinase specificities accurately identifies specific Tyr kinases that are dysregulated in cells after stimulation with growth factors, treatment with anti-cancer drugs or expression of oncogenic variants. Furthermore, the topology of known Tyr signalling networks naturally emerged from a comparison of the sequence specificities of the Tyr kinases and the SH2 phosphotyrosine (pTyr)-binding domains. Finally we show that the intrinsic substrate specificity of Tyr kinases has remained fundamentally unchanged from worms to humans, suggesting that the fidelity between Tyr kinases and their protein substrate sequences has been maintained across hundreds of millions of years of evolution.

Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 38720073 · DOI · PMC11136658

Targeted protein degradation via intramolecular bivalent glues [2]

Research article Hsia O, Hinterndorfer M, Cowan AD, Iso K, Ishida T, Sundaramoorthy R, et al. · · 2024 · cited 168 times (Europe PMC)

Abstract reading summary — source excerpts
Opening excerpt
Targeted protein degradation is a pharmacological modality that is based on the induced proximity of an E3 ubiquitin ligase and a target protein to promote target ubiquitination and proteasomal degradation.
Closing excerpt
We thus introduce a new modality in targeted protein degradation, which works by bridging protein domains in cis to enhance surface complementarity with E3 ligases for productive ubiquitination and degradation.
Interpretation limits
Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (198 words)

Targeted protein degradation is a pharmacological modality that is based on the induced proximity of an E3 ubiquitin ligase and a target protein to promote target ubiquitination and proteasomal degradation. This has been achieved either via proteolysis-targeting chimeras (PROTACs)-bifunctional compounds composed of two separate moieties that individually bind the target and E3 ligase, or via molecular glues that monovalently bind either the ligase or the target1-4. Here, using orthogonal genetic screening, biophysical characterization and structural reconstitution, we investigate the mechanism of action of bifunctional degraders of BRD2 and BRD4, termed intramolecular bivalent glues (IBGs), and find that instead of connecting target and ligase in trans as PROTACs do, they simultaneously engage and connect two adjacent domains of the target protein in cis. This conformational change 'glues' BRD4 to the E3 ligases DCAF11 or DCAF16, leveraging intrinsic target-ligase affinities that do not translate to BRD4 degradation in the absence of compound. Structural insights into the ternary BRD4-IBG1-DCAF16 complex guided the rational design of improved degraders of low picomolar potency. We thus introduce a new modality in targeted protein degradation, which works by bridging protein domains in cis to enhance surface complementarity with E3 ligases for productive ubiquitination and degradation.

Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 38383787 · DOI · PMC10917667

Sequence-specific intramembrane proteolysis: identification of a recognition motif in rhomboid substrates [3]

Research article Strisovsky K, Sharpe HJ, Freeman M · · 2009 · cited 166 times (Europe PMC)

Abstract reading summary — source excerpts
Opening excerpt
Members of the widespread rhomboid family of intramembrane proteases cleave transmembrane domain (TMD) proteins to regulate processes as diverse as EGF receptor signaling, mitochondrial dynamics, and invasion by apicomplexan parasites.
Closing excerpt
Our work demonstrates that intramembrane proteases can be sequence specific and that genome-wide substrate prediction based on their recognition motifs is feasible.
Interpretation limits
Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (139 words)

Members of the widespread rhomboid family of intramembrane proteases cleave transmembrane domain (TMD) proteins to regulate processes as diverse as EGF receptor signaling, mitochondrial dynamics, and invasion by apicomplexan parasites. However, lack of information about their substrates means that the biological role of most rhomboids remains obscure. Knowledge of how rhomboids recognize their substrates would illuminate their mechanism and might also allow substrate prediction. Previous work has suggested that rhomboid substrates are specified by helical instability in their TMD. Here we demonstrate that rhomboids instead primarily recognize a specific sequence surrounding the cleavage site. This recognition motif is necessary for substrate cleavage, it determines the cleavage site, and it is more strictly required than TM helix-destabilizing residues. Our work demonstrates that intramembrane proteases can be sequence specific and that genome-wide substrate prediction based on their recognition motifs is feasible.

Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 20064469 · DOI · PMC2941825

Key reviews and meta-analyses

The most cited review-type papers among PubMed’s best-match results for this heading, listed with bibliographic details only. Follow the links for the full text and access terms.

  • Research article Enzyme promiscuity: mechanism and applications [4]
    Hult K, Berglund P · Trends Biotechnol · 2007 · cited 328 times (Europe PMC)
  • Research article Caspase substrates [5]
    Timmer JC, Salvesen GS · Cell Death Differ · 2007 · cited 325 times (Europe PMC)
  • Research article Substrate specificity of protein kinases and computational prediction of substrates [6]
    Kobe B, Kampmann T, Forwood JK, Listwan P, Brinkworth RI · Biochim Biophys Acta · 2005 · cited 77 times (Europe PMC)
  • Research article Substrate recognition by nonribosomal peptide synthetase multi-enzymes [7]
    Lautru S, Challis GL · Microbiology (Reading) · 2004 · cited 73 times (Europe PMC)
  • Research article Molecular basis of substrate specificity in family 1 glycoside hydrolases [8]
    Marana SR · IUBMB Life · 2006 · cited 59 times (Europe PMC)
  • Research article Structural biology of the aldo-keto reductase family of enzymes: catalysis and cofactor binding [9]
    Sanli G, Dudley JI, Blaber M · Cell Biochem Biophys · 2003 · cited 57 times (Europe PMC)

References

  1. Yaron-Barir TM, Joughin BA, Huntsman EM, Kerelsky A, Cizin DM, Cohen BM, et al.. The intrinsic substrate specificity of the human tyrosine kinome. . 2024. PMID 38720073 · DOI · PMC11136658
  2. Hsia O, Hinterndorfer M, Cowan AD, Iso K, Ishida T, Sundaramoorthy R, et al.. Targeted protein degradation via intramolecular bivalent glues. . 2024. PMID 38383787 · DOI · PMC10917667
  3. Strisovsky K, Sharpe HJ, Freeman M. Sequence-specific intramembrane proteolysis: identification of a recognition motif in rhomboid substrates. . 2009. PMID 20064469 · DOI · PMC2941825
  4. Hult K, Berglund P. Enzyme promiscuity: mechanism and applications. Trends Biotechnol. 2007. PMID 17379338 · DOI
  5. Timmer JC, Salvesen GS. Caspase substrates. Cell Death Differ. 2007. PMID 17082814 · DOI
  6. Kobe B, Kampmann T, Forwood JK, Listwan P, Brinkworth RI. Substrate specificity of protein kinases and computational prediction of substrates. Biochim Biophys Acta. 2005. PMID 16172032 · DOI
  7. Lautru S, Challis GL. Substrate recognition by nonribosomal peptide synthetase multi-enzymes. Microbiology (Reading). 2004. PMID 15184549 · DOI
  8. Marana SR. Molecular basis of substrate specificity in family 1 glycoside hydrolases. IUBMB Life. 2006. PMID 16611572 · DOI
  9. Sanli G, Dudley JI, Blaber M. Structural biology of the aldo-keto reductase family of enzymes: catalysis and cofactor binding. Cell Biochem Biophys. 2003. PMID 12663943 · DOI

About this article

Sources
NLM Medical Subject Headings (definition, synonyms); NCBI PubMed (counts, identifiers); Europe PMC (open-access abstracts and citation counts).
Verification
Every reference was resolved against PubMed on 2026-10-31; title, year and DOI matched.
Review state
automated source verified; no human expert review has been recorded yet.
Version
1, last updated 2026-10-29.

See how EvidenceLib is built. Spotted an error? Email support@evidencehelix.com.

Evidence strength

EvidenceHelix's computed view of the study designs behind this topic: how common meta-analyses, randomized trials and reviews are, how recent the literature is, and a plain-language reading. It describes the design mix of the literature. It is not a quality appraisal of any study, and it is not medical advice.

Included with the Researcher plan

Reading every article stays free. This computed view is part of the paid plans.

See plans Sign in

Your notes

Notes and highlights stay in this browser only; they are not sent to EvidenceHelix.

Related research topics

  • Antibody SpecificityImmunology
  • Sensitivity and SpecificityClinical Research
  • AdiposityMolecular Biology
  • Alternative SplicingMolecular Biology
  • Amino Acid SequenceMolecular Biology
  • AnaphaseMolecular Biology

All 100 articles in Molecular Biology →

At a glance

103,969
PubMed records on this heading
8%
published since 2020
6,895
review articles
63
randomized trials
9
verified references below

Counts come from PubMed on 2026-10-29; they show how much has been written, not how strong the evidence is.

Related topics See all →

  • Antibody SpecificityImmunology
  • Sensitivity and SpecificityClinical Research
  • AdiposityMolecular Biology
  • Alternative SplicingMolecular Biology
  • Amino Acid SequenceMolecular Biology

My notes & annotations (0)

Nothing yet. Select text and press Highlight, or write a note in the Notes tab. Stored in this browser only.

Sources

  • NLM MeSH record ↗
  • PubMed search used ↗

EvidenceLib is a research-education resource, not medical advice. It does not diagnose, prescribe or recommend starting, stopping or changing any treatment. Talk to a qualified clinician about your own situation. Methods, sources and licences · Legal · Support